CN118440811A - A PCR automated all-in-one machine - Google Patents

Abstract

The present invention relates to a PCR automated all-in-one machine, comprising a frame, and a nucleic acid amplification module and a nucleic acid detection module arranged in the frame, wherein the nucleic acid amplification module is used to amplify reaction samples, and the nucleic acid detection module is used to detect reaction samples. A test kit carrying a reaction sample is placed in the nucleic acid amplification module for amplification, and after amplification, the nucleic acid detection module is detected, and the detection process is simple. A test kit can enter the PCR automated all-in-one machine for detection, and there is no need to collect a large number of samples for unified detection, and the test results are quickly given. The nucleic acid amplification module and the nucleic acid detection module occupy space, thereby improving the compactness of the PCR automated all-in-one structure and reducing the space occupied by the equipment. The configuration, transfer, amplification and detection of the reaction sample can all be completed automatically, so there is no need for professional test personnel to operate, the operation is simple, and the professional requirements for PCR detection are low. And each module is arranged in a casing, so as to avoid pollution caused by the escape of aerosols, etc.

Description

A PCR automated all-in-one machine

Technical Field

The present invention relates to the technical field of in vitro diagnosis, and in particular to an automated all-in-one PCR machine.

Background technique

PCR refers to a molecular biology experimental method for in vitro enzymatic synthesis of specific DNA fragments, which is mainly composed of repeated thermal cycles of three steps: high temperature denaturation, low temperature annealing and suitable temperature extension. Before PCR amplification, the reaction sample needs to be placed in a carrier, where the reaction sample consists of samples such as collected throat swabs or nasal swabs and reagents for PCR amplification. During PCR amplification, the reaction sample needs to be heated by a heater and cooled by a cooling mechanism, so that the reaction sample cycles through high temperature denaturation, low temperature annealing and suitable temperature extension stages.

The PCR detection equipment in the prior art requires professional testers to operate to form reaction samples, the operation is complicated, and the PCR detection has high professional requirements.

To avoid aerosol contamination, the equipment needs to be placed in a special room, and PCR testing equipment has high requirements for the environment.

The PCR testing equipment is not compact and takes up a lot of space. In addition, a large number of samples need to be collected and tested uniformly, and the test results cannot be given quickly.

Summary of the invention

The purpose of the present invention is to provide a PCR automated all-in-one machine to solve one of the above technical problems.

To achieve the above objectives, the present invention provides a PCR automated all-in-one machine, comprising a frame, and a nucleic acid amplification module and a nucleic acid detection module arranged on the frame, wherein the nucleic acid amplification module is used to amplify reaction samples, and the nucleic acid detection module is used to detect reaction samples.

Optionally, the PCR automated all-in-one machine further includes a preparation module, which is disposed on the rack and is used to prepare samples and reagents into reaction samples.

Optionally, the preparation module includes a carrying submodule and a pipetting submodule arranged on the upper side of the carrying submodule, and the carrying submodule and the pipetting submodule can move relative to each other so that the pipetting submodule transfers the solution on the carrying submodule and injects the reaction sample into the reagent kit.

Optionally, the pipetting submodule includes a pipetting gun, and at least one of the pipetting gun and the carrying submodule can move in a horizontal plane, and at least one of the pipetting gun and the carrying submodule can move in a vertical direction.

Optionally, the carrying submodule is capable of carrying a reagent kit, and the pipetting submodule is used to transfer solutions between cavities of the reagent kit.

Optionally, the carrying submodule includes a bracket, and a receiving portion is provided on the bracket, and the receiving portion is used to receive the reagent kit.

Optionally, the receiving portion includes a plurality of receiving grooves, into which the reagent carrying portion of the reagent kit can be inserted.

Optionally, a heating structure is provided in at least one of the plurality of receiving grooves, and the heating structure is used to heat the reaction sample in the reagent carrying part.

Optionally, the PCR automated all-in-one machine further includes a transfer module disposed on the frame, and the transfer module is used to transfer the test kit or carrier between the carrying submodule and the nucleic acid amplification module, or between the carrying submodule, the nucleic acid amplification module and the nucleic acid detection module.

Optionally, the PCR automated all-in-one machine further comprises a cutting module disposed on the frame, wherein the cutting module is used to cut the test kit between the carrier of the test kit and the reagent carrying portion of the test kit.

Optionally, the PCR automated integrated machine further comprises a flip module disposed on the frame, and the flip module is used to flip the carrier of the reagent kit to a preset angle.

Optionally, the PCR automated all-in-one machine further comprises a bending module disposed on the frame, the bending module being used to bend one end of the reagent kit at a preset angle, or the bending module being used to bend the carrier of the reagent kit at a preset angle and break the reagent kit between its carrier and the reagent carrying portion.

Optionally, the reaction sample is contained in a carrier, the carrier is a flat structure, and the nucleic acid amplification module and/or the nucleic acid detection module enables the carrier to be vertically inserted.

Optionally, the PCR automated all-in-one machine further includes a control module, and the control module is used to control the nucleic acid amplification module and the nucleic acid detection module.

Optionally, the nucleic acid detection module includes a fluorescence detection optical path system, and the fluorescence detection optical path system includes:

At least one fluorescent light emitting unit, configured to emit excitation light.

Optionally, the number of the fluorescence emission units is at least two, and the control module controls at least two of the fluorescence emission units to emit excitation light in different time periods.

Optionally, the fluorescence detection optical path system further includes:

The fluorescence detection unit comprises at least two fluorescence transmission light paths, and the at least two fluorescence emission units correspond one to one with the at least two fluorescence transmission light paths.

Optionally, the fluorescence detection unit further includes a detector, and the plurality of fluorescence transmission light paths are all connected to the detector.

Optionally, the number of the detector is one, and the detector is electrically connected to the control module to record the intensity of the fluorescence signal in different time periods.

Optionally, the fluorescence detection optical path system also includes a fiber optic holder, the fluorescence transmission optical path includes a collecting optical fiber, an output end of the excitation optical fiber that emits the excitation light, and an input end of the collecting optical fiber that injects the fluorescence signal form a fiber group, the fiber group is arranged in the fiber optic holder, and the optical fibers in the fiber optic holder are arranged along a preset direction, which is the radial direction of the optical fiber, so that the output end of the collecting optical fiber and the input end of the excitation optical fiber are arranged flat in the fiber optic holder.

Optionally, at least the two fluorescence transmission light paths and the at least two fluorescence emission units form at least two groups of optical fiber groups, and the at least two groups of optical fiber groups are arranged in sequence along the preset direction.

Optionally, one of the optical fiber groups includes at least two of the collecting optical fibers, and at least one of the collecting optical fibers is disposed on both sides of the preset direction of the excitation optical fiber in one of the optical fiber groups.

Optionally, the nucleic acid amplification module includes at least one set of temperature regulating mechanisms, and the temperature regulating mechanisms are capable of changing the temperature of the reaction sample in the carrier.

Optionally, the nucleic acid amplification module includes two groups of temperature regulation mechanisms, which are respectively a first temperature regulation mechanism and a second temperature regulation mechanism. The positions of the first temperature regulation mechanism and/or the second temperature regulation mechanism are adjustable so as to be closer to or farther away from each other, and so that the carrier is clamped between the first temperature regulation mechanism and the second temperature regulation mechanism.

Optionally, the nucleic acid amplification module includes a driving mechanism, and the driving mechanism drives the first temperature regulating mechanism and/or the second temperature regulating mechanism to move closer to or away from each other.

Optionally, a plurality of groups of elastic leveling components are arranged at intervals between the driving mechanism and the first temperature regulating mechanism and/or the second temperature regulating mechanism.

Optionally, the nucleic acid amplification module further includes an extrusion mechanism, and the first temperature regulating mechanism and/or the second temperature regulating mechanism are connected to the extrusion mechanism to extrude an extrusion cavity of a carrier clamped between the first temperature regulating mechanism and the second temperature regulating mechanism.

Optionally, a step mounting hole is provided on the first temperature regulating mechanism and/or the second temperature regulating mechanism, and the extrusion mechanism includes an extrusion assembly and a buffer member, the extrusion assembly is connected to the step mounting hole, and the buffer member is connected between the extrusion assembly and the step mounting hole so that the extrusion assembly is in elastic contact with the carrier.

Optionally, both the first temperature regulating mechanism and the second temperature regulating mechanism include a cooling component for cooling the carrier.

Optionally, the first temperature regulating mechanism and the second temperature regulating mechanism both further include a heater, and the heater is arranged on a side of the first temperature regulating mechanism and the second temperature regulating mechanism where the cooling components are close to each other.

Optionally, the heater includes a heating element, and the PCR automated all-in-one machine further includes a resistance detection element, and the resistance detection element is used to detect the resistance of the heating element.

Optionally, the PCR automated all-in-one machine further includes a first temperature detection unit for detecting the temperature of the heater.

Optionally, the nucleic acid amplification module further includes a positioning mechanism for positioning the carrier, and the positioning mechanism is connected to the first temperature regulating mechanism or the second temperature regulating mechanism.

Optionally, the flat structure refers to a structure in which the ratio of the dimension of the carrier in a direction perpendicular to its thickness direction to the dimension of the carrier in its thickness direction is greater than 5:1.

Optionally, the ratio of the dimensions is 50:1 to 100:1.

As can be seen from the above, the technical solution provided by the present invention is to place the reagent kit carrying the reaction sample into the nucleic acid amplification module for amplification, and then detect it in the nucleic acid detection module after amplification, and the detection process is simple. A reagent kit can enter the PCR automated all-in-one machine for detection, and there is no need to collect a large number of samples for unified detection, and the test results are quickly given. The nucleic acid amplification module and the nucleic acid detection module take up little space, thereby improving the compactness of the structure of the PCR automated all-in-one machine and reducing the space occupied by the equipment. According to the time period, it is determined what kind of signal fluorescence is, the equipment cost is low, the equipment structure is simple, there is no mechanical switching, and the detection speed is fast. The configuration, transfer, amplification and detection of the reaction sample can all be completed automatically, so it is simple to operate without professional test personnel, and the PCR detection professional requirements are low. And each module is arranged in the casing to avoid the escape of aerosols, etc., so even if the PCR automated all-in-one machine is not placed in a special room, it will not cause pollution.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1a is a front view of a PCR automated integrated machine provided in an embodiment of the present invention;

FIG1b is a three-dimensional diagram of an automated integrated PCR machine provided in an embodiment of the present invention;

FIG2a is a schematic diagram of the structure of a kit provided in an embodiment of the present invention;

FIG2b is a cross-sectional view of a carrier provided by an embodiment of the present invention;

FIG2c is a schematic structural diagram of a carrier from another perspective provided by an embodiment of the present invention;

FIG3 is a schematic diagram of the structure of a partial PCR automated integrated machine provided in an embodiment of the present invention;

FIG4 is a schematic diagram of a process of preparing reaction samples by an automated integrated PCR machine provided in an embodiment of the present invention;

FIG5a is a schematic structural diagram of a bracket provided in an embodiment of the present invention;

FIG5b is a schematic structural diagram of another bracket provided by an embodiment of the present invention;

FIG6a is a schematic diagram of the structure of a vector provided by an embodiment of the present invention when it is inserted into a nucleic acid amplification module;

FIG6b is another schematic diagram of the structure of a vector provided by an embodiment of the present invention when it is inserted into a nucleic acid amplification module;

FIG7a is a schematic structural diagram of another cutting module provided by an embodiment of the present invention;

FIG7 b is a three-dimensional diagram of another PCR automated integrated machine provided by an embodiment of the present invention;

FIG7c is a partial enlarged view of point B in FIG7b;

FIG8a is a schematic structural diagram of a first bending module provided by an embodiment of the present invention;

FIG8b is a schematic structural diagram of a second bending module provided by an embodiment of the present invention;

FIG8c is a front view of another PCR automated integrated machine provided by an embodiment of the present invention;

9 is a schematic diagram of the structure of a carrier, a nucleic acid amplification module and a nucleic acid detection module provided in an embodiment of the present invention;

10 is a schematic structural diagram of a nucleic acid detection module and a nucleic acid amplification module (the first temperature adjustment mechanism and the second temperature adjustment mechanism do not clamp the carrier) provided in an embodiment of the present invention;

11 is a top view of a nucleic acid detection module and a nucleic acid amplification module in a normal use state provided by an embodiment of the present invention;

FIG12 is a schematic diagram of the structure of another nucleic acid amplification module provided in an embodiment of the present invention;

FIG13 is an exploded view of the nucleic acid amplification module in FIG12 ;

FIG14 is a cross-sectional view of the nucleic acid amplification module in FIG12 at a first position;

FIG15 is a schematic structural diagram of a cooling body provided by an embodiment of the present invention;

FIG16 is a cross-sectional view of the second position of the nucleic acid amplification module in FIG12;

FIG17 is a cross-sectional view of the nucleic acid amplification module in FIG12 at a third position;

FIG18 is a cross-sectional view of the nucleic acid amplification module in FIG12 at a fourth position;

FIG19 is a partial enlarged view of point A in FIG18;

FIG20 is a schematic diagram of the structure of another nucleic acid amplification module provided in an embodiment of the present invention;

FIG21 is a schematic structural diagram of another nucleic acid amplification module provided by an embodiment of the present invention from another perspective;

22 is a schematic structural diagram of a carrier, a carrying plate and a positioning mechanism provided in an embodiment of the present invention;

23 is a schematic structural diagram of a carrier plate and a positioning mechanism provided in an embodiment of the present invention;

FIG24a is a schematic diagram of a partial structure of a nucleic acid amplification module provided by an embodiment of the present invention;

FIG24b is a schematic diagram of the structure of a carrier plate provided in an embodiment of the present invention;

FIG25 is a temperature-time curve of a carrier provided in an embodiment of the present invention;

26 is a schematic diagram of the structure of a heater provided in an embodiment of the present invention;

FIG27 is a schematic diagram of the structure of a nucleic acid detection module and a carrier provided in an embodiment of the present invention;

FIG28 is a schematic diagram of the structure of a fluorescence detection optical path in the prior art;

FIG29 is a schematic structural diagram of a fluorescence detection unit provided by the present invention;

FIG30 is a schematic diagram of the structure of another fluorescence detection unit provided in an embodiment of the present invention;

31 is a schematic diagram of the structure of the optical fiber holder provided in an embodiment of the present invention;

32 is a schematic structural diagram of an optical fiber holder from another perspective according to an embodiment of the present invention;

FIG33 is a schematic diagram of the structure of a nucleic acid detection module provided by an embodiment of the present invention for detecting from one side of a carrier;

Figure 34 is a structural schematic diagram of the nucleic acid detection module provided by an embodiment of the present invention for detecting from both sides of the carrier.

In the figure:

1. Fluorescence emission unit; 11. Excitation optical fiber; 12. Light source;

2. Fluorescence detection unit; 21. Fluorescence transmission optical path; 211. Filter; 212. Collecting optical fiber; 213. Collimating lens; 22. Detector; 23. Turntable;

4. carrier; 41. amplification chamber; 42. side wall; 43. first wall; 44. second wall; 45. bending notch; 46. extrusion chamber;

51. elastic leveling assembly; 511. connecting rod; 512. elastic member; 513. first flange;

52. A first temperature regulating mechanism;

521, cooling assembly; 5211, cooling body; 52111, cooling shell; 52112, heat sink; 52113, cooling groove; 52116, medium port; 52117, temperature measuring hole; 52118, temperature measuring protrusion; 5212, base; 5213, electrical connection line; 5215, insulation layer; 5216, second temperature detection unit; 5217, cover; 5218, first sealing ring; 5219, second sealing ring;

522, heater; 523, step mounting hole; 5231, first step surface; 5232, second step surface; 524, adapter plate;

53. Driving mechanism; 531. Driving member; 532. Mounting frame; 533. Screw rod; 534. Nut; 535. Driving gear; 536. Driven gear; 537. Bearing;

54. guide mechanism; 541. guide rail; 542. slider;

55. Support seat;

56. second temperature adjustment mechanism; 561. carrier plate; 5611. through hole;

57, positioning mechanism; 5711, first positioning portion; 5712, second positioning portion; 5713, first plate; 5714, second plate; 572, second positioning assembly; 5721, fixing portion; 5722, elastic adjustment portion; 5723, abutting portion; 5724, chamfer;

58, extrusion mechanism; 581, buffer; 582, mounting rod; 583, pressure head; 584, second flange;

91. Heating element; 92. Upper conducting component; 921. Heat-saturating layer; 922. Insulating layer; 93. Temperature calibration unit; 94. Fast conducting unit; 941. SMD; 942. Guide pillar; 95. Lower conducting component; 951. Insulating thermal resistance layer; 952. Thermal conductive layer; 96. Wiring port; 97. External electrical connection contact; 98. Electrical connection lead; 99. First temperature detection unit;

10. Nucleic acid amplification module;

20. Nucleic acid detection module; 201. Fluorescence detection optical path system; 2011. Optical fiber group; 2012. Optical fiber holder;

30. preparation module; 31. bearing submodule; 311. bracket; 3111. receiving slot; 3112. heating structure; 3113. slot wall;

32. Liquid transfer submodule; 321. Liquid transfer gun; 322. Vertical drive member; 323. Horizontal drive member; 324. Liquid transfer connector; 325. Liquid transfer gear; 326. Rack;

40. Rack;

50. Control module;

60. Transfer module; 61. Pick-and-place member; 66. Vertical transfer drive member; 67. Horizontal transfer drive member; 68. Transfer gear; 69. Transfer connector;

70, cutting module; 701, cutting drive assembly; 702, first cutter; 703, first cutter;

80, bending module; 801, pressure column; 802, horizontal driving member; 803, vertical driving member; 804, second roller; 805, first roller; 806, handle; 807, pressure column lifting driving member; 808, bending driving structure;

90. Recycling module;

7. Flip module; 71. Flip driving member; 72. First plug-in member; 73. Second plug-in member;

100, reagent box; 1001, reagent carrying part; 1011, preset reagent chamber; 1012, injection chamber; 1013, cavity; 1014, sealing film;

300. Nucleic acid amplification detection device.

In Figure 28:

200, excitation light path; 201, fluorescence signal light path; 203, dichroic mirror; 204, filter; 205, light source; 206, detection circuit.

Detailed ways

The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. It is understood that the specific embodiments described herein are only used to explain the present invention, rather than to limit the present invention. It is also necessary to explain that, for ease of description, only the parts related to the present invention are shown in the accompanying drawings, rather than all.

Some directional words are defined in the present invention. Unless otherwise specified, the directional words used, such as "up", "down", "left", "right", "inside" and "outside", are used to facilitate understanding and therefore do not constitute a limitation on the scope of protection of the present invention.

In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

In the description of the present invention, unless otherwise clearly specified and limited, the terms "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be a direct connection or an indirect connection through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

The technical solution of the present invention is further described below in conjunction with the accompanying drawings and through specific implementation methods. It is understood that the specific embodiments described herein are only used to explain the present invention, rather than to limit the present invention. It is also necessary to explain that, for ease of description, only the parts related to the present invention are shown in the accompanying drawings, rather than all.

Some directional words are defined in the present invention. Unless otherwise specified, the directional words used, such as "up", "down", "left", "right", "inside" and "outside", are used to facilitate understanding and therefore do not constitute a limitation on the scope of protection of the present invention.

In the present invention, unless otherwise clearly specified and limited, a first feature being "above" or "below" a second feature may include that the first and second features are in direct contact, or may include that the first and second features are not in direct contact but are in contact through another feature between them. Moreover, a first feature being "above", "above" and "above" a second feature includes that the first feature is directly above and obliquely above the second feature, or simply indicates that the first feature is higher in level than the second feature. A first feature being "below", "below" and "below" a second feature includes that the first feature is directly below and obliquely below the second feature, or simply indicates that the first feature is lower in level than the second feature.

In the description of the present invention, unless otherwise clearly specified and limited, the terms "connected", "connected", and "fixed" should be understood in a broad sense, for example, it can be a fixed connection, a detachable connection, or an integral body; it can be a mechanical connection or an electrical connection; it can be directly connected or indirectly connected through an intermediate medium, it can be the internal connection of two elements or the interaction relationship between two elements. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

As shown in Fig. 1a and Fig. 1b, the PCR automated integrated machine provided in this embodiment includes a nucleic acid amplification detection device 300, which includes a nucleic acid amplification module 10 and a nucleic acid detection module 20. The nucleic acid amplification module 10 is used to amplify the reaction sample, and the nucleic acid detection module 20 is used to detect the reaction sample. The PCR automated integrated machine may also include a rack 40, and the nucleic acid amplification module 10 and the nucleic acid detection module 20 are both arranged on the rack 40.

The reagent kit 100 carrying the reaction sample is placed in the nucleic acid amplification module 10 for amplification, and then detected in the nucleic acid detection module 20 after amplification. The detection process is simple.

As shown in Figures 1a and 1b, illustratively, there can be two nucleic acid amplification detection devices 300, and the nucleic acid amplification modules 10 and the nucleic acid detection modules 20 are set one by one, and the number is equal. Of course, the number of nucleic acid amplification detection devices 300 is not limited to two, and can also be one or more. After the nucleic acid amplification module 10 realizes the amplification of the reaction sample, it continues to clamp the carrier 4 so that the nucleic acid detection module 20 detects the reaction sample.

In other optional embodiments, as shown in FIG3 , the nucleic acid amplification module 10 and the nucleic acid detection module 20 are separately arranged. After the nucleic acid amplification module 10 realizes the amplification of the reaction sample, the carrier 4 of the nucleic acid amplification module 10 needs to be transferred to the nucleic acid detection module 20 so that the nucleic acid detection module 20 detects the reaction sample. The number of nucleic acid amplification modules 10 or nucleic acid detection modules 20 may be different, and the number may be one or more. Preferably, the number of nucleic acid amplification modules 10 and nucleic acid detection modules 20 may be arranged proportionally according to the time to complete the nucleic acid amplification and nucleic acid detection of the reaction sample in a single carrier 4. The number of nucleic acid amplification modules 10 and nucleic acid detection modules 20 is coordinated, and the ratio of the number of nucleic acid amplification modules 10 to the number of nucleic acid detection modules 20 may be not less than 1, so as to avoid the utilization of the nucleic acid amplification modules 10 and the nucleic acid detection modules 20 being sufficient. If it takes 8 minutes for the nucleic acid amplification module 10 to complete an amplification and 2 minutes for the nucleic acid detection module 20 to complete a detection, the number of nucleic acid amplification modules 10 may be 3-4 times that of the nucleic acid detection modules 20.

Optionally, the PCR automated all-in-one machine may further include a housing (not shown in the figure), and the nucleic acid amplification module 10 and the nucleic acid detection module 20 are both arranged in the housing to prevent the escape of aerosols, etc., so that even if the PCR automated all-in-one machine is not placed in a special room, it will not cause pollution.

As shown in Figure 2a, the present disclosure also provides a reagent kit 100 for use with a PCR automated all-in-one machine, and the reagent kit 100 is a consumable, that is, disposable. The reagent kit 100 includes a reagent carrying part 1001 and a carrier 4, and the reagent carrying part 1001 is at least used to carry reagents, wherein the reagents include but are not limited to sample treatment liquid, PCR reaction buffer and enzyme system. Carrier 4 is used to carry reaction samples. It is understandable that the reaction sample is formed by mixing samples such as collected throat swabs or nasal swabs and reagents used in the reagent carrying part 1001. After the sample is collected, the reagents in the reagent carrying part 1001 can be directly used to configure the reaction sample, thereby improving the detection efficiency. At the same time, the reagents in the reagent carrying part 1001 can be quantitatively placed according to the actual amount required, and can be operated without professional test personnel, which has universal applicability. In addition, there is no need for the PCR equipment to specifically set up containers for carrying reagents and configuring reagents, and there is no need to repeatedly replenish reagents in the PCR automated all-in-one machine, thereby simplifying the structure and operation of the PCR automated all-in-one machine.

The reagent carrying portion 1001 may include at least one preset reagent chamber 1011, in which the reagent is placed. The reagent carrying portion 101 may also include at least one injection chamber 1012 and/or at least one cavity 1013. The injection chamber 1012 may be used to place a pharyngeal swab or a liquid sample, and the cavity 1013 may be used to mix reagents. A total of 5 injection chambers 1012, preset reagent chambers 1011, and cavities 1013 may be provided. Of course, more than 5 or less than 5 may be provided as needed. In an optional embodiment, the injection chamber 1012 is used to accommodate a sample processing liquid, and samples such as pharyngeal swabs or nasal swabs collected by the user may be placed in the injection chamber 1012. Two preset reagent chambers 1011 are provided, and are used to accommodate a PCR reaction buffer and an enzyme system, respectively.

At least one injection cavity 1012, at least one preset reagent cavity 1011, and at least one cavity 1013 are provided with openings for reagents or samples to enter and exit the cavity. A sealing film 1014 is provided on the opening. Before placing the sample into the reagent box 100, the sealing film 1014 is opened. Before that, the cavity is isolated from the outside world, which can ensure the cleanliness of the cavity.

As shown in Fig. 2a to Fig. 2c, the carrier 4 includes a first wall 43 and a second wall 44 arranged opposite to each other, and a side wall 42 arranged between the first wall 43 and the second wall 44. The first wall 43, the second wall 44 and the side wall 42 form an amplification chamber 41. The carrier 4 is a flat structure, and at least part of the side wall 42 is light-transmissive. In particular, fluorescence can pass through the light-transmissive side wall 42 of the amplification chamber 41, so that the nucleic acid detection module 20 can be detected through the side wall 42. Preferably, the amplification chamber 41 is also a flat structure.

It can be understood that the flat structure may refer to that the dimension of the carrier 4 or the amplification chamber 41 in the thickness direction (i.e., the direction in which the first wall 43 and the second wall 44 are arranged) is much smaller than the dimension in the direction perpendicular to the thickness direction. As an example, the ratio of the dimension in the direction perpendicular to the thickness direction to the dimension in the thickness direction is greater than 5:1, such as the dimension ratio is 50:1 to 100:1. As an example, the dimension ratio is 90:1. As an example, the amplification chamber 41 is a cuboid, and the ratio of the length and thickness of the cuboid may be greater than 5:1, such as 90:1. For example, the dimension in the thickness direction of the amplification chamber 41 may be 0.3-1.0 mm, and the width and length of the amplification chamber 41 may be about 10 mm and 20 mm, respectively. As an example, the amplification chamber 41 may also be a cylindrical structure, and the ratio of the diameter to the thickness is greater than 5:1, such as the thickness is 0.3-1.0 mm and the diameter is 5-20 mm. Of course, the cross section of the amplification chamber 41 may be polygonal or elliptical, etc.

Optionally, the first wall 43 and the second wall 44 of the carrier 4 are made of a heat-conducting material, such as an aluminum film, or an aluminum film and an isolation film, and optionally, the isolation film is a polypropylene film (i.e., PP film). The isolation film is in direct contact with the reaction sample, which can prevent the aluminum film from affecting the reaction sample, thereby facilitating rapid heat conduction between the reaction sample in the carrier 4 and the temperature regulating mechanism.

As shown in FIG. 2c, as a preferred embodiment, the carrier 4 may further include an extrusion chamber 46, which is connected to the amplification chamber 1. The extrusion chamber 46 is deformed under the external force F so that the extrusion chamber 46 squeezes the amplification chamber 41, and the extrusion chamber 46 controls the pressure in the amplification chamber 41. For example, during PCR amplification, the extrusion chamber 46 is squeezed, and the gas and/or liquid (the liquid may be a reaction sample) in the extrusion chamber 46 applies pressure to the reaction sample in the amplification chamber 41, and the reaction sample squeezes the cavity wall corresponding to the amplification chamber 41, so that the cavity wall corresponding to the amplification chamber 41 expands outward, so that the outer side of the cavity wall corresponding to the amplification chamber 41 is in full contact with the nucleic acid amplification module 10. At the same time, the reaction sample is attached to the inner side of the cavity wall of the amplification chamber 41 under the pressure of the extrusion chamber 46, and because the outer side of the cavity wall of the amplification chamber 41 is in full contact with the nucleic acid amplification module 10, the amplification efficiency can be greatly improved.

As shown in Fig. 1a and Fig. 1b, optionally, the PCR automated integrated machine may further include a preparation module 30, which is disposed in the frame 40. The preparation module 30 is used to prepare the sample and the reagent into a reaction sample. Specifically, the preparation module 30 mixes the reagent in the reagent carrying part 1001 with the sample to form a reaction sample, and injects the reaction sample into the carrier 4. As an example, the preparation module 30 is disposed in the housing.

As shown in FIG. 1a , the preparation module 30 includes a carrying submodule 31 and a liquid transfer submodule 32 disposed on the upper side of the carrying submodule 31. The carrying submodule 31 and the liquid transfer submodule 32 can move relative to each other, so that the liquid transfer submodule 32 transfers the solution on the carrying submodule 31, mixes the sample with the reagent to form a reaction sample, and the carrying submodule 31 and the liquid transfer submodule 32 can move relative to each other, and can also inject the reaction sample in the reagent carrying part 1001 into the carrier 4. The carrying submodule 31 and the liquid transfer submodule 32 cooperate with each other to complete the configuration of the reaction sample and place the reaction sample in the carrier 4 to prepare for nucleic acid amplification.

As shown in Fig. 1b, specifically, the carrying submodule 31 is used to carry the reagent box 100, and the pipetting submodule 32 is used to transfer the solution between the cavities of the reagent box 100. That is, the pipetting submodule 32 can absorb the sample, reagent and reaction sample in the reagent box 100 and transfer the sample, reagent and reaction sample.

Optionally, the pipetting submodule 32 includes a pipetting gun 321, which can absorb reagents and discharge reagents into the reagent box 100. At least one of the pipetting gun 321 and the carrying submodule 31 can move in a horizontal plane, and at least one of the pipetting gun 321 and the carrying submodule 31 can move in a vertical direction.

Exemplarily, the carrying submodule 31 is located below the pipetting submodule 32. The carrier 4, the injection chamber 1012, the preset reagent chamber 1011 and the cavity 1013 are arranged along the first direction (the X direction as shown in Figures 1a-2), and the pipette gun 321 of the pipetting submodule 32 reciprocates along the first direction and the vertical direction (the Z direction as shown in Figures 1 and 2) relative to the frame 40 to achieve liquid preparation. Among them, the first direction is perpendicular to the vertical direction, and the first direction is the horizontal direction.

The liquid transfer submodule 32 further includes a vertical drive member 322 and a horizontal drive assembly, wherein the vertical drive member 322 is connected to the liquid transfer gun 321 to drive the liquid transfer gun 321 to reciprocate in the vertical direction, and the horizontal drive assembly is connected to the vertical drive member 322 to drive the vertical drive member 322 to reciprocate in the first direction, thereby driving the liquid transfer gun 321 to reciprocate in the first direction. The horizontal drive assembly and the vertical drive member 322 may be connected via a liquid transfer connector 324. Optionally, the vertical drive member 322 may be a drive member such as a cylinder or an electric cylinder that can drive the liquid transfer gun 321 to reciprocate in the vertical direction.

Exemplarily, the horizontal driving assembly includes a horizontal driving member 322, a liquid transfer gear 325 and a rack 326. The horizontal driving member 322 can be a servo motor, etc. The liquid transfer gear 325 is connected to the output end of the horizontal driving member 322. The liquid transfer gear 325 is meshed with the rack 326 for transmission. The rack 326 is connected to the frame 40 and extends along the first direction. The horizontal driving member 322 such as the servo motor rotates forward and reverse, thereby driving the liquid transfer gear 325 to rotate forward and reverse. The liquid transfer gear 325 is meshed with the rack 326 for transmission, so that the horizontal driving member 322 reciprocates along the first direction, thereby driving the vertical driving member 322 and the liquid transfer gun 321 to reciprocate along the first direction.

As shown in Figure 1b, when the pipette gun 321 needs to absorb the reagent in the reagent box 100, the vertical drive member 322 drives the pipette gun 321 to move downward and extends into the reagent to absorb the reagent. After the absorption is completed, the vertical drive member 322 drives the pipette gun 321 to move upward and is located above the reagent box 100. The horizontal drive component drives the reagent box 100 to translate, so that the pipette gun 321 is vertically aligned with the cavity that needs to be injected. The vertical drive member 322 drives the pipette gun 321 to move downward to inject the reagent into another cavity of the reagent box 100.

FIG4 schematically shows the process of using a PCR automated all-in-one machine to configure a reaction sample and inject the reaction sample into a carrier 4:

Step 1: tear off the sealing film 1014 outside the PCR automated integrated machine manually or with automated equipment;

Step 2: manually or automatically placing the collected sample into the injection chamber 1012 outside the PCR automated all-in-one machine;

Step 3, the reagent kit 100 is placed in the carrying submodule 31, the vertical driving member 322 and the horizontal driving assembly drive the liquid transfer gun 321 to move, and the liquid transfer gun 321 transfers the PCR reaction buffer in one of the preset reagent chambers 1011 to the cavity 1013;

Step 4, the vertical driving member 322 and the horizontal driving assembly drive the liquid transfer gun 321 to move, and the liquid transfer gun 321 transfers the enzyme system in another preset reagent chamber 1011 to the cavity 1013;

Step 5, the vertical driving member 322 and the horizontal driving assembly drive the liquid transfer gun 321 to move, and the liquid transfer gun 321 transfers the sample processing liquid in the injection chamber 1012 to the cavity 1013;

Step 6, mixing the reaction sample in the cavity 1013 by using the pipette gun 321;

Step 7, the vertical driving member 322 and the horizontal driving assembly drive the liquid transfer gun 321 to move, and the liquid transfer gun 321 transfers the reaction sample in the cavity 1013 to the carrier 4;

Step 8, complete the injection.

As shown in Figures 1a and 1b, optionally, the PCR automated all-in-one machine also includes a transfer module 60 arranged on the frame 40, and the transfer module 60 is used to transfer the test kit 100 or the carrier 4 between the carrying submodule 31 and the nucleic acid amplification module 10, or as shown in Figure 3, the transfer module 60 is used to transfer the test kit 100 or the carrier 4 between the carrying submodule 31, the nucleic acid amplification module 10 and the nucleic acid detection module 20. As an example, the transfer module 60 is arranged above the nucleic acid amplification module 10 and the carrying submodule 31. The configuration, transfer, amplification and detection of the reaction sample can all be completed automatically, so there is no need for professional testers to operate the operation, and the PCR detection professional requirements are low. As an example, the transfer module 60 is arranged in the casing, and preferably, each module is arranged in the casing, and an air purification module can be arranged in the casing to avoid aerosols and the like from escaping and causing pollution.

As shown in Fig. 1a and Fig. 1b, illustratively, the transfer module 60 provided by the present disclosure includes a horizontal transfer drive assembly, a vertical transfer drive member 66 and a pick-and-place member 61, wherein the pick-and-place member 61 is used to grab and put down the carrier 4, the reagent carrying portion 1001 or the reagent reagent 100, the vertical transfer drive member 66 is connected to the pick-and-place member 61 to drive the pick-and-place member 61 to reciprocate in the vertical direction, and the horizontal transfer drive assembly is connected to the vertical transfer drive member 66 to drive the vertical transfer drive member 66 to reciprocate in the first direction, thereby driving the pick-and-place member 61 to reciprocate in the first direction. Among them, the horizontal transfer drive assembly and the vertical transfer drive member 66 can be connected by a transfer connector 69. Optionally, the vertical transfer drive member 66 can be a drive member such as a cylinder or an electric cylinder that can drive the pick-and-place member 61 to reciprocate in the vertical direction.

The pick-and-place member 61 can transfer the carrier 4, the reagent carrying part 1001 or the reagent reagent box 100 by vacuum adsorption or clamping. As an example, when the pick-and-place member 61 transfers the carrier 4, the reagent carrying part 1001 or the reagent reagent box 100 by clamping, the pick-and-place member 61 can be a pneumatic finger as shown in Figures 1a and 1b, and when the pick-and-place member 61 transfers the carrier 4, the reagent carrying part 1001 or the reagent reagent box 100 by vacuum adsorption, the pick-and-place member 61 can include a suction cup connected to a vacuum pump, etc., to generate a vacuum environment between the suction cup and the adsorption reagent box 100 or the carrier 4, thereby adsorbing the reagent carrying part 1001, the reagent reagent box 100 or the carrier 4.

Exemplarily, the horizontal transfer drive assembly includes a horizontal transfer drive member 67, a transfer gear 68 and a transfer rack. The horizontal transfer drive member 67 can be a servo motor, etc. The transfer gear 68 is connected to the output end of the horizontal transfer drive member 67, and the transfer gear 68 is meshed with the transfer rack for transmission. Optionally, the transfer rack is the above-mentioned rack 326, that is, the pipetting submodule 32 and the transfer module 60 share the same rack 326, that is, as shown in Figure 1b, the horizontal drive assembly and the horizontal transfer drive assembly are connected to the rack 326 along the first direction, and the pipetting gear 325 of the horizontal drive assembly and the transfer gear 68 of the horizontal transfer drive assembly are respectively meshed with the rack 326. When the transfer module 60 picks up the carrier 4 located on the carrying submodule 31, the pipetting submodule 32 can be located at one end of the rack 326, and the above-mentioned one end is the end of the rack 326 close to the carrying submodule 31, that is, the position shown in Figure 1b, so as to avoid the transfer module 60, thereby avoiding interference with the transfer module 60. The horizontal transfer drive member 67 such as the servo motor rotates forward and reverse, thereby driving the transfer gear 68 to rotate forward and reverse. The transfer gear 68 meshes with the rack 326 for transmission, so that the horizontal transfer drive member 67 moves back and forth along the first direction, thereby driving the vertical transfer drive member 66 and the pick-and-place member 61 to move back and forth along the first direction.

The following describes the working process of the transfer module 60 by taking the transfer module 60 transferring the carrier 4 from the carrier module 31 to the nucleic acid amplification module 10 as an example:

The horizontal transfer drive assembly drives the pick-and-place member 61 to be located directly above the carrier 4 on the carrying submodule 31;

The vertical transfer drive 66 drives the pick-and-place member 61 to move downward to pick up the carrier 4;

The vertical transfer drive 66 drives the pick-and-place member 61 to move upward to prevent the carrier 4 from colliding with other structures during the transfer of the carrier 4;

The horizontal transfer drive assembly drives the pick-and-place member 61 to move toward the side where the nucleic acid amplification module 10 is located, until it moves to the top of the nucleic acid amplification module 10;

The vertical transfer drive 66 drives the pick-and-place member 61 to move downward to insert the carrier 4 into the nucleic acid amplification module 10 .

It is understandable that the carrier 4 corresponding to this transfer module 60 is placed vertically when placed in the housing, so the carrier 4 does not need to be rotated, and the transfer module 60 only drives the carrier 4 to translate and can be vertically inserted into the nucleic acid amplification module 10. As shown in Figure 7a, the thickness direction of the carrier 4 is perpendicular to the depth direction of the cavity of the reagent carrying part 1001, so as to ensure that the carrier 4 is not rotated, and the carrier 4 can also be vertically inserted into the nucleic acid detection module 10, while ensuring that the opening of the cavity of the reagent carrying part 1001 is facing upward. Of course, the carrier 4 and the reagent carrying part 1001 can also be set separately, in which case the transfer module 60 only drives the carrier 4 to translate and inserts the carrier 4 vertically into the nucleic acid amplification module 10.

As shown in FIG. 1a and FIG. 1b , optionally, the PCR automated all-in-one machine further includes a recovery module 90 disposed in the housing and used for recovering the reagent kit 100 . The recovery module 90 is connected to the frame 40 and is located below the transfer module 60 .

The transfer module 60 can transfer the reagent kit 100 in the housing to the recovery module 90. Specifically, the transfer module 60 can grab the reagent kit 100, the reagent carrying part 1001 or the carrier 4 after detection, and transfer them to the recovery module 90.

As shown in FIG. 1 , illustratively, the carrier module 31 , the nucleic acid amplification detection device 300 and the recovery module 90 are all arranged in sequence along the first direction to facilitate the arrangement of the transfer module 60 and improve the compactness of the PCR automated all-in-one structure.

As shown in Fig. 3 and Fig. 5a, in order to stably place the reagent box 100, the carrying submodule 31 optionally includes a bracket 311, and a receiving portion is provided on the bracket 311, and the receiving portion is used to receive the reagent box 100. On the one hand, the receiving portion can limit the position of the reagent box 100 to ensure that each reagent box 100 is placed at a fixed position on the carrying submodule 31; on the other hand, during the entire pipetting process, the movement of the reagent box 100 relative to the bracket 311 can be limited to ensure that the pipette gun 321 can accurately enter the cavity of the reagent box 100.

Optionally, the receiving portion includes a plurality of receiving grooves 3111, and the reagent carrying portion 1001 of the reagent kit 100 can be at least partially inserted into the receiving grooves 3111. Further, the receiving grooves 3111 correspond to the respective cavities of the reagent kit 100, and the respective cavities are respectively inserted into the receiving grooves 3111, and the groove walls 3113 between two adjacent receiving grooves 3111 can support the reagent kit 100 and improve the stability of the reagent kit 100.

A heating structure 3112 is provided in at least one of the multiple receiving grooves 3111. The heating structure 3112 can heat the reagents in the reagent kit 100 in advance so that the reagents are at a suitable temperature in advance, thereby reducing the time required for the reaction sample to be amplified in the nucleic acid amplification module 10. For example, the heating structure 3112 can heat the reagents to 25°C-80°C±1°C. It can be understood that the heating structure 3112 can be a heating element such as a heating wire. As long as the heating structure 3112 can be built into the receiving groove 3111 and heat the solution, the heating structure 3112 is a prior art, so it will not be described here. At the same time, the receiving groove 3111 corresponds to each cavity of the reagent kit 100, and the ambient temperature of the cavity of the reagent kit 100 can also be controlled separately.

As shown in Fig. 5a, the bracket 311 can support the carrier 4 and the reagent carrying part 1001, wherein one of the receiving grooves 3111 can accommodate the carrier 4, and the carrier 4 can be placed horizontally on the bracket 311, that is, the thickness direction of the carrier 4 is consistent with the vertical direction. As shown in Fig. 5b, the bracket 311 can also be used only to support the reagent carrying part 1001 without supporting the carrier 4, and the carrier 4 is placed in the air.

In the present embodiment, preferably, the nucleic acid amplification module 10 and/or the nucleic acid detection module 20 make the carrier 4 be inserted vertically, that is, the thickness direction of the carrier 4 is perpendicular to the vertical direction. When performing PCR amplification and/or detection, the carrier 4 is placed vertically, that is, the carrier 4 is inserted into the nucleic acid amplification module 10 and/or the nucleic acid detection module 20 in the direction shown by the arrows in Figures 6a and 6b, and the thickness direction of the carrier 4 (the X direction as shown in Figures 6a and 6b) is perpendicular to the vertical direction (the Z direction as shown in Figures 6a and 6b, the three directions of X, Y and Z are perpendicular to each other). In this way, when the carrier 4 is inserted into the nucleic acid amplification module 10 and/or the nucleic acid detection module 20, the bubble can float to the top of the carrier 4, avoiding the bubble from being dispersed in various places of the side wall 42 of the carrier 4, thereby ensuring the accuracy of the nucleic acid detection module 20 detection, and avoiding the problem of uneven temperature of the reaction sample. During the heating process, the gas in the reaction sample is precipitated to generate bubbles. When the carrier 4 is inserted vertically, the bubble can flow upward to the top of the carrier 4, thereby not affecting the detection and amplification. At the same time, convection is generated by the gravity of the reaction sample itself, and the movement of bubbles disturbs the reaction sample, which can further increase the consistency of the reaction sample temperature.

7a Using the PCR automated all-in-one machine provided in this embodiment, one reagent kit 100 can enter the PCR automated all-in-one machine for testing, without the need to collect a large number of samples for unified testing, and the test results can be given quickly.

As shown in FIG1b , optionally, the PCR automated all-in-one machine further includes a cutting module 70 (a first cutter 703 as shown in FIG1b ), and the cutting module 70 is used to cut the reagent kit 100 between the carrier 4 and the reagent carrying part 1001. The cutting module 70 can cut the reagent kit 100 before the transfer module 60 transfers the carrier 4 to the nucleic acid amplification module 10. After the reagent kit 100 is cut, the transfer module 60 can only transfer the carrier 4 to the nucleic acid amplification module 10 and the nucleic acid detection module 20, so that the reagent carrying part 1001 can be prevented from occupying space in the nucleic acid amplification module 10 and the nucleic acid detection module 20, thereby improving the compactness of the structure of the PCR automated all-in-one machine and reducing the space occupied by the equipment.

Of course, in other optional embodiments, the carrier 4 and the reagent carrying portion 1001 may be separately provided, so that the reagent kit 100 does not need to be cut and a cutting module 70 does not need to be provided.

As shown in FIG. 1b , the first cutting module 70 is exemplarily disclosed to include a first cutter 703. A storage slot can be provided on the carrier module 31. The first cutter 703 can be placed in the storage slot or taken out from the storage slot. After the first cutter 703 is placed in the storage slot, one end of the first cutter 703 extends out of the storage slot. The pick-and-place member 61 of the transfer module 60 (at this time, the pick-and-place member 61 transfers the carrier 4 by clamping and placing so as to clamp the first cutter 703) clamps the end of the first cutter 703 extending out of the storage slot to cut the reagent kit 100.

Exemplarily, the process of cutting the reagent kit 100 may be as follows:

The horizontal transfer drive assembly drives the pick-and-place member 61 to be located directly above the first cutter 703;

The vertical transfer drive 66 drives the pick-and-place member 61 to move downward to pick up the first cutter 703;

The vertical transfer drive 66 drives the pick-and-place member 61 to move upward to take the first cutter 703 out of the storage slot;

The horizontal transfer drive assembly drives the pick-and-place member 61 to move toward the side where the reagent kit 100 is located, until it moves to just above the position where the reagent kit 100 needs to be cut;

The vertical transfer drive 66 drives the pick-and-place member 61 to move downward to cut the reagent kit 100;

The horizontal transfer drive assembly drives the pick-and-place member 61 to drive the first cutter 703 to move toward the side where the storage slot is located, until it moves to the top of the storage slot;

The vertical transfer driving member 66 drives the pick-and-place member 61 to move downward to place the first cutter 703 into the storage slot.

As shown in Figure 7a, a second cutting module 70 is exemplarily disclosed, and the cutting module 70 includes a cutting drive component 701 and a second cutter 702. The cutting drive component 701 is used to drive the second cutter 702 to approach and cut the reagent kit 100, and drive the second cutter 702 to avoid the reagent kit 100, so that the pipette gun 321 can transfer liquid, and the transfer module 60 can transfer the reagent kit 100.

Specifically, the cutting drive assembly 701 includes a cutting rotation drive member and a crank-connecting rod slider mechanism. The cutting rotation drive member can be a motor, etc., to drive one end of the crank-connecting rod slider mechanism to rotate. The second cutter 702 is connected to the other end of the crank-connecting rod slider mechanism to realize the up and down movement of the second cutter 702.

Optionally, the cutting drive assembly 701 can be connected to the frame 4 through a driving structure (not shown in the figure), and the driving structure drives the cutting drive assembly 701 to move along the first direction, so as to avoid the transfer module 60 and the pipetting submodule 32 when the transfer module 60 and the pipetting submodule 32 move along the first direction. The driving structure can be the same as the structure of the horizontal driving assembly and the horizontal transfer driving assembly, and share the rack 326 with the horizontal driving assembly and the horizontal transfer driving assembly.

As shown in Figures 7b and 7c, optionally, the PCR automated integrated machine further includes a flip module 7 disposed on the frame, and the flip module 7 is used to flip the carrier of the test kit to a preset angle. When the carrier 4 is placed in the carrying submodule 31, when the thickness direction of the carrier 4 is parallel to the vertical direction, the carrier 4 can be flipped 90° by the flip module 7, so that the thickness direction of the carrier 4 is perpendicular to the vertical direction. It is understandable that the reagent carrying portion 1001 and the carrier 4 of the test kit 100 can be a split structure, and the flip module 7 only flips the carrier 4 90°, or the reagent carrying portion 1001 and the carrier 4 of the test kit 100 are an integrated structure, and the reagent carrying portion 1001 and the carrier 4 are separated by the cutting module 70, so that the flip module 7 only flips the carrier 4 90°.

As shown in Fig. 7c, the flip module 7 can be arranged on the bearing submodule 31, and the flip module 7 can be located on one side of the first cutter 703. The flip module 7 can include a flip driver 71 and a first connector 72. The flip driver 71 can be a stepping motor, etc. The output end of the flip driver 71 is connected to the first connector 72 for driving the first connector 72 to flip. The first connector 72 can include a first slot, and a portion of the carrier 4 can be plugged into the slot. The flip driver 71 drives the first connector 72 to flip 90°, and the carrier 4 in the first slot flips 90° accordingly. When the first connector 72 is not flipped, the first slot can extend in the horizontal direction to facilitate the insertion of the carrier 4, and when the carrier 4 is flipped, the carrier 4 is prevented from being separated from the first slot.

Optionally, the flip module 7 may further include a second connector 73, the second connector 73 may include a second slot, and the reagent carrying part 1001 may be disposed in the second slot. The second slot may extend in a horizontal direction to facilitate the insertion of the reagent carrying part 1001.

The flip module 7 provided in this embodiment has a simple structure and is easy to operate, and can improve the compactness of the PCR automated all-in-one machine.

As shown in Fig. 1b and Fig. 8a, in other optional embodiments, the flip module 7 may not be provided, but the PCR automated integrated machine further includes a bending module 80 provided on the frame 40, the bending module 80 being used to bend one end of the reagent box 100 at a preset angle, specifically, the bending module 80 bends the carrier 4 of the reagent box 100 at a preset angle, such as 90°. As shown in Fig. 8a, the bending module 80 bends the horizontally placed carrier 4 to a vertical position. In order to facilitate the bending of the carrier 4, optionally, a bending notch 45 is provided at the connection between the carrier 4 and the reagent carrying portion 1001.

Optionally, the reagent kit 100 is disposed on the carrying submodule 31, and the end of the reagent kit 100 to be bent is suspended. In this embodiment, the end to be bent is the carrier 4. The reagent carrying part 1001 can be placed on the bracket 311 as shown in FIG. 5b, and the bracket 311 can make the carrier 4 suspended.

As shown in FIG. 1b and FIG. 8a, the first bending module 80 is exemplarily disclosed to include a first roller 805 and a handle 806 connected to each other. A storage slot can be provided on the carrying submodule 31. The first roller 805 and the handle 806 can be placed in the storage slot or taken out from the storage slot. After the bending module 80 is placed in the storage slot, one end of the handle 806 extends out of the storage slot. The pick-and-place member 61 of the transfer module 60 picks up and places one end of the handle 806 extending out of the storage slot to bend the reagent kit 100.

Exemplarily, the process of bending the reagent kit 100 may be as follows:

The horizontal transfer drive assembly drives the pick-and-place member 61 to be located directly above the handle 806;

The vertical transfer drive 66 drives the pick-and-place member 61 to move downward to pick up the bending module 80; (Since the vertical transfer drive 66 can be a cylinder or an electric cylinder, when the vertical transfer drive 66 drives the pick-and-place member 61 to be located at a higher position, the lower end of the vertical transfer drive 66 is also located at a higher position. Therefore, when the horizontal transfer drive assembly drives the vertical transfer drive 66 to move along the first direction, it will not interfere with other modules.)

The vertical transfer drive 66 drives the pick-and-place member 61 to move upward to take the bending module 80 out of the storage tank;

The horizontal transfer drive assembly drives the pick-and-place member 61 to move toward the side where the reagent kit 100 is located, until the first roller 805 moves to the top of the carrier 4;

The vertical transfer driving member 66 drives the pick-and-place member 61 to move downward until the first roller 805 abuts against the carrier 4;

Bending the reagent kit 100: the vertical transfer drive 66 drives the pick-and-place member 61 to continue to move downward (since the vertical transfer drive 66 can be a pneumatic cylinder or an electric cylinder, etc., it can drive the pick-and-place member 61 to move downward, thereby driving the bending module 80 to move downward), and at the same time, the horizontal transfer drive assembly drives the pick-and-place member 61 toward the reagent carrying portion 1001, so that the second roller 804 moves in a circular arc trajectory to always press against the surface of the carrier 4, thereby pushing the carrier 4 to bend. The second roller 804 can reduce the friction between it and the surface of the carrier 4, thereby protecting the carrier 4 from being scratched. As shown in Figure 8a, the position of the carrier 4 in the initial state, the intermediate state and the final state are shown. During the bending process, the pipette gun 321 can be made to abut against the reagent carrying portion 1001 to prevent the reagent carrying portion 1001 from tilting upward;

Put the bending module 80 back into the storage tank: the horizontal transfer drive assembly drives the pick-and-place member 61 to move the bending module 80 to the side where the storage tank is located, until it moves to just above the storage tank; the vertical transfer drive member 66 drives the pick-and-place member 61 to move downward to place the bending module 80 into the storage tank;

The pick-and-place member 61 of the transfer module 60 returns and picks up the carrier 4 to transfer the carrier 4 to the nucleic acid amplification module 10 .

As shown in FIG. 8 b , optionally, the present disclosure further provides a structural schematic diagram of a second bending module 80 , in which the bending module 80 is movably connected to the frame 40 along a first direction.

The bending module 80 includes a dual-axis module, the lower end of which can move along the first direction and the vertical direction to always press against the upper surface of the carrier 4 and bend the carrier 4 at a preset angle. As shown in FIG8b , the positions of the carrier 4 in the initial state, the intermediate state and the final state are shown.

Optionally, the dual-axis module includes a transverse drive member 802, a vertical drive member 803 and a second roller 804, wherein the vertical drive member 803 is connected to the output end of the transverse drive member 805, and the second roller 804 is connected to the output end of the vertical drive member 803. The transverse drive member 802 is used to drive the vertical drive member 803 and the second roller 804 to move along the first direction, that is, to realize the movement of the bending module 80 relative to the frame 40 along the first direction. The transverse drive member 802 can have the same structure as the horizontal drive assembly and the horizontal transfer drive assembly, and can also share the rack 326 with the horizontal drive assembly and the horizontal transfer drive assembly. In this case, the transverse drive member 802 is connected to the frame 40 through the rack 326, and the transverse drive member 802, the horizontal drive assembly and the horizontal transfer drive assembly are arranged on the frame 40 at intervals along the first direction. The vertical drive member 803 is used to drive the second roller 804 to move along the vertical direction, and the structure of the vertical drive member 803 can be the same as that of the vertical transfer drive member 66. The second roller 804 can press against the upper surface of the carrier 4 .

In the process of bending the carrier 4, the lateral drive member 802 cooperates with the vertical drive member 803 so that the second roller 804 moves in an arc trajectory to always press against the surface of the carrier 4, thereby pushing the carrier 4 to bend. The second roller 804 can reduce the friction between it and the surface of the carrier 4, thereby protecting the carrier 4 from being scratched. On the other hand, as shown in Figures 8b and 8c, the lateral drive member 802 drives the vertical drive member 803 and the second roller 804 to move along the first direction, so as to avoid the transfer module 60 and the transfer submodule 32 when the transfer module 60 and the transfer submodule 32 move along the first direction. Exemplarily, the right end of the rack 326 shown in Figure 8c extends out of the right end of the bearing submodule 31, and the bending module 80 and the transfer submodule 32 can move to the right end of the rack 326 to avoid the transfer module 60, thereby avoiding interference with the transfer module 60.

Optionally, the bending module 80 may further include a pressure column 801, which can be pressed against the upper surface of the reagent carrying part 1001, and the pressure column 801 may be connected to a pressure column lifting drive component 807 such as a cylinder, and the pressure column lifting drive component 807 may drive the pressure column 801 to reciprocate in the vertical direction to press against the carrier 4, thereby preventing the reagent carrying part 1001 from tilting upward during the bending of the carrier 4. The pressure column lifting drive component 807 may also be connected to a bending drive structure 808, and the bending drive structure 808 drives the pressure column lifting drive component 807 and the pressure column 801 to move in the first direction, so as to avoid the transfer module 60 and the pipetting submodule 32 when the transfer module 60 and the pipetting submodule 32 move in the first direction. The bending drive structure 808 may have the same structure as the horizontal drive assembly and the horizontal transfer drive assembly, and may also share the rack 326 with the horizontal drive assembly and the horizontal transfer drive assembly.

Of course, the bending module 80 may not include the pressure column 801, the pressure column lifting and lowering drive component 807 and the bending drive structure 808. Instead, like the first bending module 80, the reagent carrying part 1001 is prevented from tilting upward by making the pipette gun 321 abut against the reagent carrying part 1001.

In other optional embodiments, the bending module 80 bends the carrier 4 of the reagent kit 100 at a preset angle, such as bending 90° and breaking the reagent kit 100, such as breaking the reagent kit 100 between the carrier 4 and the reagent carrying portion 1001. Exemplarily, at least the connection between the carrier 4 and the reagent carrying portion 1001 is made of a brittle material, such as the brittle material may be acrylic (i.e. PMMA), polystyrene (i.e. PS), etc., so that the reagent kit 100 is broken after the reagent kit 100 is bent. The connection between the carrier 4 and the reagent carrying portion 1001 may be provided with a breaking opening such as a stamp hole that makes it easy for the reagent kit 100 to be broken. In addition, the carrying submodule 31 may be provided with a guide groove that is adapted to the contour of the carrier 4 so that the carrier 4 can be kept in a vertical state when it is bent to a vertical state.

As shown in Figure 1b and Figure 9 (in order to facilitate the display of the structure of the nucleic acid amplification module 10, the placement direction of the nucleic acid amplification module 10 in Figure 9 is not the placement direction when it is used), in this embodiment, the nucleic acid amplification module 10 includes at least one set of temperature regulation mechanisms, which can change the temperature of the reaction sample in the carrier 4 so that the temperature of the reaction sample is repeatedly thermally cycled in three stages of high temperature denaturation, low temperature annealing and suitable temperature extension.

Exemplarily, the nucleic acid amplification module 10 includes two sets of temperature regulating mechanisms, as shown in FIG9 , the two sets of temperature regulating mechanisms can be close to or away from each other, so that the carrier 4 is sandwiched between the two sets of temperature regulating mechanisms. Specifically, the two sets of temperature regulating mechanisms are respectively a first temperature regulating mechanism 52 and a second temperature regulating mechanism 56, and the position of the first temperature regulating mechanism 52 and/or the second temperature regulating mechanism 56 is adjustable to be close to or away from the second temperature regulating mechanism 56, and the carrier 4 is sandwiched between the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56. For example, the first temperature regulating mechanism 52 abuts against one of the first wall 43 and the second wall 44 of the carrier 4, and the second temperature regulating mechanism 56 abuts against the other of the first wall 43 and the second wall 44 of the carrier 4, thereby clamping the carrier 4, and the side wall 42 of the carrier 4 is exposed for detection by the nucleic acid detection module 20.

As shown in FIG. 1b, the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are arranged along the first direction, and the nucleic acid detection module 20 can be arranged on one side or both sides of the second direction (the Y direction as shown in FIG. 1b) of the nucleic acid amplification module 10, so that when the nucleic acid amplification module 10 clamps the carrier 4, the nucleic acid detection module 20 can detect the reaction sample in the carrier 4 through the side wall 42 of the carrier 4. Among them, the first direction, the second direction and the vertical direction are perpendicular to each other. Of course, in other optional embodiments, the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are arranged along the second direction, and the nucleic acid detection module 20 can be arranged on one side or both sides of the first direction of the nucleic acid amplification module 10.

Specifically, as shown in Figure 10 (to facilitate the display of the structure of the nucleic acid amplification module 10, the placement direction of the nucleic acid amplification module 10 in Figure 10 is not the placement direction when it is in use), when the first temperature adjustment mechanism 52 is away from the second temperature adjustment mechanism 56, the carrier 4 can be taken out from between the first temperature adjustment mechanism 52 and the second temperature adjustment mechanism 56, or the carrier 4 can be placed between the first temperature adjustment mechanism 52 and the second temperature adjustment mechanism 56.

As shown in Figures 11 and 9, when the first temperature adjustment mechanism 52 is close to the second temperature adjustment mechanism 56, the first temperature adjustment mechanism 52 and the second temperature adjustment mechanism 56 can clamp the carrier 4, thereby heating or cooling the carrier 4. At this time, the fluorescence detection optical path system 201 performs fluorescence detection through the side wall 42 of the carrier 4, and the optical fiber seat 2012 is set on one side of the carrier 4 to fix the optical fiber, so that the optical fiber passes through the side wall 42 of the carrier 4 for fluorescence detection.

As shown in FIG. 11 , a top view of the nucleic acid detection module 200 and the nucleic acid amplification module 100 in a normal use state is shown, and the nucleic acid detection module 200 and the nucleic acid amplification module 100 are placed in such a way that the carrier 4 is placed vertically after being inserted between the first temperature adjustment mechanism 52 and the second temperature adjustment mechanism 56. At this time, the bearing submodule 31 can be arranged on one side of the nucleic acid detection module 200 and the nucleic acid amplification module 100 as shown in FIG. 11 (in FIG. 11 , the bearing submodule 31 is arranged on the upper side of the nucleic acid detection module 200 and the nucleic acid amplification module 100, and in other optional embodiments, the bearing submodule 31 can be arranged on the lower side of the nucleic acid detection module 200 and the nucleic acid amplification module 100, and it can be understood that the upper side and the lower side are both directions in the figure, not the directions in actual use).

As shown in Fig. 9, the nucleic acid amplification module 10 includes a driving mechanism 53, and the driving mechanism 53 drives the first temperature regulating mechanism 52 and/or the second temperature regulating mechanism 56 to move closer to or away from each other. The following is an exemplary description using the example of the driving mechanism 53 driving the first temperature regulating mechanism 52 to move along the first direction and move closer to or away from the second temperature regulating mechanism 56.

As shown in FIGS. 12 and 13 , optionally, the nucleic acid amplification module 10 may include a support base 55 , and the driving mechanism 53 and the second temperature adjustment mechanism 56 may be disposed on the support base 55 .

As shown in FIG12 , in order to make the first temperature adjustment mechanism 52 move stably, the nucleic acid amplification module 10 may further include a guide mechanism 54 for guiding the temperature adjustment mechanism. The guide mechanism 54 may be disposed on a support seat 55 , and the support seat 55 may be connected to the frame 40 .

Specifically, the guide mechanism 54 includes a guide rail 541 and a slider 542. The guide rail 541 is installed on the support seat 55. The slider 542 is slidably connected to the guide rail 541. The first temperature adjustment mechanism 52 is connected to the slider 542. The slider 542 slides along the guide rail 541, thereby driving the first temperature adjustment mechanism 52 to slide.

As shown in FIG. 12 and FIG. 13 , the driving mechanism 53 includes a driving component and a mounting frame 532 connected to the output end of the driving component. Specifically, the first temperature adjustment mechanism 52 is connected to the slider 542 via the mounting frame 532 .

As shown in FIG. 13 , and in combination with FIG. 12 and FIG. 14 , the driving assembly may include a driving member 531 such as a transmission assembly and a servo motor, and the transmission assembly may include a driving gear 535 (as shown in FIG. 18 ), a belt (not shown in the figure), a driven gear 536, a screw 533 and a nut 534, wherein the driving member 531 may be fixed to a support seat 55 (as shown in FIG. 12 ), and the screw 533 is rotatably connected to the support seat 55 through a bearing 537. The output end of the driving member 531 may be connected to the driving gear 535, and one end of the screw 533 is connected to the driven gear 536, and the driven gear 536 and the driving gear 535 are connected through a belt transmission. The nut 534 is threadedly connected to the screw 533, and the mounting bracket 532 is connected to the nut 534.

The driving member 531 drives the driving gear 535 to rotate, and the driving gear 535 drives the belt to rotate, so that the belt drives the driven gear 536 to rotate. Since the driven gear 536 is connected to the screw rod 533, it can drive the screw rod 533 to rotate. The nut 534 is threadedly connected to the screw rod 533, so that the nut 534 moves along the direction in which the screw rod 533 extends, and then drives the mounting frame 532 to move, and the mounting frame 532 drives the first temperature regulating mechanism 52 to move. Optionally, the screw rod 533 extends along the first direction, and the extension direction of the screw rod 533 is consistent with the direction in which the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are arranged in sequence, so that when the nut 534 moves along the screw rod 533, it drives the first temperature regulating mechanism 52 to approach or move away from the second temperature regulating mechanism 56.

Of course, in other optional embodiments, the driving mechanism 53 can also simultaneously drive the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 to move closer to or away from each other. In this case, the screw 533 can be a forward and reverse thread bidirectional screw 533, and both sets of temperature regulating mechanisms are connected to mounting frames 532, and the mounting frames 532 are respectively connected to nuts 534, the nuts 534 connected to the first temperature regulating mechanism 52 are connected to the forward threads of the screw 533, and the nuts 534 connected to the second temperature regulating mechanism 56 are connected to the reverse threads of the screw 533, and then when the driving member 531 drives the screw 533 to rotate, the nuts 534 on the forward threads and the nuts 534 on the reverse threads move toward each other, so that the driving mechanism 53 simultaneously drives the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 to move closer to or away from each other.

As shown in FIG. 13 and FIG. 14 , the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 both include a cooling component 521 for cooling the carrier 4 . Optionally, the cooling components 521 of the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are symmetrically arranged to cool both sides of the carrier 4 simultaneously.

Optionally, the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 both further include a heater 522, which is disposed on a side of the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 where the cooling components 521 are close to each other, and the heater 522 is used to heat the carrier 4. In this embodiment, one side of the heater 522 is in direct contact with the carrier 4, and the side of the heater 522 away from the carrier 4 is in direct contact with the cooling component 521, that is, the cooling component 521, the heater 522 and the carrier 4 are in contact in sequence, and when the cooling component 521 cools the carrier 4, it first cools the heater 522 and then cools the carrier 4.

The nucleic acid amplification module 10 provided in this embodiment includes the following steps:

The carrier 4 is sandwiched between the first temperature regulating mechanism 52 and the second temperature regulating mechanism 51;

The cooling component 521 maintains continuous cooling, so that the cooling component 521 continuously cools the reaction sample;

During the heating process, the power of the heater 522 is increased to heat the reaction sample to the denaturation temperature or the extension temperature;

During the cooling process, the power of the heater 522 is reduced to cool the reaction sample to the annealing temperature.

Or include the following steps:

The carrier 4 is sandwiched between the first temperature regulating mechanism 52 and the second temperature regulating mechanism 51;

The cooling component 521 maintains continuous cooling, so that the cooling component 521 continuously cools the reaction sample;

Increasing the power of the heater 522 to raise the temperature of the reaction sample to the denaturation temperature;

Controlling the power of the heater 522 to keep the reaction sample at the denaturation temperature for a first preset time;

During the cooling process, the power of the heater 522 is reduced, and the reaction sample is cooled to the annealing temperature;

In the low-temperature annealing stage, the power of the heater 522 is adjusted to keep the reaction sample at the annealing temperature for a second preset time;

The above steps are cycled multiple times until a preset cycle number or a preset amplification level is reached.

During the heating process, the power of the heater 522 is adjusted to heat the reaction sample to the denaturation temperature or the extension temperature.

The reaction sample is kept continuously refrigerated by the cooling assembly 521, so that the part of the heater 522 close to the cooling assembly 521 is always maintained at a relatively low temperature. When the heat of the reaction sample is transferred through the heater 522 to achieve the heating or cooling of the reaction sample, since the part of the heater 522 close to the cooling assembly 521 is always maintained at a relatively low temperature, when the reaction sample needs to be cooled, the cooling assembly 521 only needs to cool the other part of the heater 522 and the reaction sample. That is, since the volume of the heater 522 that needs to be cooled is reduced, the entire volume that needs to be cooled is reduced, thereby shortening the required time for cooling.

During the cooling, heating, low-temperature annealing stage, high-temperature denaturation stage or suitable temperature extension stage, the heater 522 is controlled so that another part of the reaction sample close to the heater 522 reaches the desired temperature. The heater 522 is also controlled so that the reaction sample can be kept warm, cooled quickly and heated quickly. The process of raising, lowering and cooling the reaction sample is controlled within 2.5 seconds, thereby greatly shortening the detection time.

In addition, the cooling component 521 continues to cool, and before the reaction sample needs to be cooled, the cooling component 521 has been cooled in advance and has cooled the heater 522, so that it can seamlessly connect with the cooling demand of the reaction sample, and therefore, the cooling speed can be particularly improved.

As shown in Figures 13, 14 and 15, specifically, the cooling component 521 includes a cooling body 5211 and a base 5212. The cooling liquid can flow through the cooling body 5211. The base 5212 is provided with a mounting groove. The cooling body 5211 is installed in the mounting groove. The heater 522 contacts the end face of the cooling body 5211.

As shown in FIG. 15 , further, the cooling body 5211 includes a heat sink 52112 , and a flow channel for the liquid cooling medium to flow is formed between two adjacent heat sinks 52112 . The cooling medium flows continuously, taking away the heat conducted from the carrier 4 and the heater 522 to the cooling body 5211 .

Optionally, the cooling body 5211 further comprises a cold shell 52111, the cold shell 52111 is provided with a cooling groove 52113, the heat sink 52112 is connected to the cold shell 52111, the cold shell 52111 and the heat sink 52112 can be integrally formed by casting or welding, and the cold shell 52111 is arranged in the cooling groove 52113. The cold shell 52111 can also be provided with a medium port 52116 connected with the cooling groove 52113, the number of the medium ports 52116 can be two, the cooling medium enters the cooling groove 52113 through one medium port 52116, and flows out of the cooling groove 52113 through another medium port 52116.

As shown in FIGS. 13 and 14, the cooling assembly 521 may further include a cover 5217, which is covered on the notch of the cooling groove 52113. A first sealing ring 5218 may be provided between the cover 5217 and the cold shell 52111 to prevent leakage of the coolant in the cooling groove 52113. As shown in FIGS. 14 and 15, optionally, the opening direction of the notch of the cooling groove 52113 is toward the bottom of the mounting groove of the base 5212, so that after the cover 5217 is covered on the notch of the cooling groove 52113, the bottom of the mounting groove of the base 5212 can support the cover 5217. One end of the cold shell 52111 away from the cover 5217 contacts the heater 522.

As shown in Figure 13, the second temperature adjustment mechanism 56 also includes a supporting plate 561, which is provided with a through hole 5611. One end of the cooling component 521 of the second temperature adjustment mechanism 56 is penetrated through the through hole 5611 (that is, the end of the cold shell 52111 away from the cover body 5217 is penetrated through the through hole 5611), so that the end surface of the cooling body 5211 extends out of the through hole 5611 (as shown in Figure 14, the end surface of the cooling body 5211 is located on the upper side of the supporting plate 561. It can be understood that the upper side, lower side, left side or right side referred to in this embodiment are relative to the drawings and do not refer to the direction in actual use), and protrude from the supporting plate 561, so that the heater 522 is placed on the end surface of the cooling body 5211 of the second temperature adjustment mechanism 56, ensuring that the cooling body 5211 can cool the heater 522.

Optionally, the cooling component 521 also includes a thermal insulation layer 5215, which is arranged on the cooling body 5211 and located on the side where the two cooling bodies 5211 are close to each other. The thermal insulation layer 5215 can separate the cooling body 5211 from the supporting plate 561, thereby avoiding heat exchange between the cooling body 5211 and the supporting plate 561, and avoiding heat loss of the cooling body 5211.

As shown in Figures 13 to 15, preferably, the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 also include a second temperature detection unit 5216. A temperature measuring hole 52117 is opened in the cooling component 521. The second temperature detection unit 5216 is arranged in the temperature measuring hole 52117 to detect the temperature of the cooling component 521. The second temperature detection unit 5216 can be electrically connected to the control module, and then the control module adjusts the temperature of the coolant entering the cooling tank 52113 according to the temperature of the cooling component 521.

As shown in FIGS. 15 and 16 , optionally, a temperature measuring protrusion 52118 is connected to the cooling groove 52113 of the cold shell 52111, a temperature measuring hole 52117 is provided on the cover 5217 and the temperature measuring protrusion 52118, and a second sealing ring 5219 is provided between the temperature measuring protrusion 52118 and the cover 5217, so as to prevent the coolant from entering the temperature measuring hole 52117, thereby protecting the second temperature detection unit 5216. Optionally, the second temperature detection unit 5216 includes a temperature sensor, and the second temperature detection unit 5216 is electrically connected to the control module 50 via an electrical connection line 5214.

As shown in FIG. 14 , optionally, multiple sets of elastic leveling components 51 are arranged between the driving mechanism 53 and the first temperature regulating mechanism 52, and the elastic leveling components 51 are elastic so as to level the first temperature regulating mechanism 52, so that the first temperature regulating mechanism 52 is in full contact with the carrier 4, and to avoid the first temperature regulating mechanism 52 from being in hard contact with the carrier 4 and damaging the carrier 4. Of course, when the driving mechanism 53 only drives the second temperature regulating mechanism 56 to move, multiple sets of elastic leveling components 51 are arranged between the driving mechanism 53 and the second temperature regulating mechanism 56; or when the driving mechanism 53 drives the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 to move, multiple sets of elastic leveling components 51 are arranged between the driving mechanism 53 and the first temperature regulating mechanism 52 and between the driving mechanism 53 and the second temperature regulating mechanism 56.

As shown in FIG. 16 , specifically, the elastic leveling component 51 is connected between the mounting frame 532 and the first temperature regulating mechanism 52, so that when the first temperature regulating mechanism 52 contacts the carrier 4, if the first temperature regulating mechanism 52 is tilted relative to the carrier 4, the first temperature regulating mechanism 52 closer to the carrier 4 contacts the carrier 4 first, and the first temperature regulating mechanism 52 that is not in contact with the carrier 4 can continue to move, and the elastic leveling component 51 corresponding to the first temperature regulating mechanism 52 that contacts the carrier 4 is compressed until the first temperature regulating mechanism 52 is completely in contact with the carrier 4. In this way, before the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 approach each other, whether the mounting frame 532 is parallel to the surface of the carrier 4, and whether the first temperature regulating mechanism 52 is parallel to the surface of the carrier 4, will not affect the temperature of the first temperature regulating mechanism 52 and the surface of the carrier 4. This reduces the accuracy requirements for the mounting frame 532, the first temperature regulating mechanism 52, and the connection between the two.

As shown in FIG. 17 , the elastic leveling assembly 51 includes a connecting rod 511 and an elastic member 512. The connecting rod 511 is slidably arranged on the mounting frame 532. One end of the connecting rod 511 is connected to the first temperature regulating mechanism 52. The elastic member 512 is sleeved on the connecting rod 511 and is located between the mounting frame 532 and the first temperature regulating mechanism 52. The elastic member 512 may be a spring, etc. When the elastic leveling assembly 51 is compressed, the connecting rod 511 slides relative to the mounting frame 532 to reduce the length of the connecting rod 511 between the first temperature regulating mechanism 52 and the mounting frame 532. At the same time, the elastic member 512 is compressed until the first temperature regulating mechanism 52 is completely in contact with the carrier 4.

Optionally, a hole is opened on the mounting frame 532, and the end of the connecting rod 511 can be connected to the first flange 513, and the first flange 513 abuts against the surface of the mounting frame 532 away from the first temperature regulating mechanism 52, thereby preventing the connecting rod 511 from falling off when the first temperature regulating mechanism 52 is away from the second temperature regulating mechanism 56.

As shown in Figure 17, an adapter plate 524 can also be set between the elastic leveling component 51 and the cooling component 521 of the first temperature adjustment mechanism 52. The cooling component 521 and the adapter plate 524 can be fixedly connected by bolts or the like. The elastic leveling component 51 and the first temperature adjustment mechanism 52 are connected via the adapter plate 524 to facilitate the connection of the elastic leveling component 51.

As shown in FIGS. 18 to 20 and 22, the nucleic acid amplification module 10 further includes an extrusion mechanism 58, and the first temperature regulating mechanism 52 and/or the second temperature regulating mechanism 56 are connected with the extrusion mechanism 58 to squeeze the extrusion cavity 46 of the carrier 4 clamped on the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56. That is, an external force F is applied to the extrusion cavity 46 by the extrusion mechanism 58, so that the extrusion cavity 46 is deformed, and then the extrusion cavity 46 controls the pressure in the amplification cavity 41.

Specifically, as shown in Figure 18, the first temperature regulating mechanism 52 is connected to the extrusion mechanism 58, and as shown in Figures 20 and 21, the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are both connected to the extrusion mechanism 58. Of course, in other optional embodiments, the second temperature regulating mechanism 56 can also be connected to the extrusion mechanism 58.

As shown in FIG18 , optionally, when the first temperature regulating mechanism 52 is connected to the extrusion mechanism 58, after the carrier 4 is placed on the second temperature regulating mechanism 56, the extrusion cavity 46 can be directly opposite to the carrier plate 561, so that the carrier plate 561 supports the extrusion cavity 46. As shown in FIG20 and FIG21 , when both the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are connected to the extrusion mechanism 58, the two extrusion mechanisms 58 are directly opposite to each other to extrude the extrusion cavity 46.

As shown in Figures 14, 18 and 22, the following description is made by taking the extrusion mechanism 58 disposed on the first temperature regulating mechanism 52 as an example. It can be understood that the connection method between the second temperature regulating mechanism 56 and the extrusion mechanism 58 is roughly the same as the connection method between the first temperature regulating mechanism 52 and the extrusion mechanism 58, so it is not repeated. Optionally, a step mounting hole 523 is provided on the base 5212 of the first temperature regulating mechanism 52, and the extrusion mechanism 58 includes an extrusion assembly and a buffer 581, the extrusion assembly is connected to the step mounting hole 523, and the buffer 581 is connected between the extrusion assembly and the step mounting hole 523, so that the extrusion assembly is in elastic contact with the carrier 4, thereby preventing the carrier 4 from being damaged. Optionally, the buffer 581 is a spring.

As shown in FIG. 19 , further, the extrusion assembly includes a mounting rod 582 and a pressing head 583, the mounting rod 582 is slidably arranged in the step mounting hole 523, and the first step surface 5231 of the step mounting hole 523 can limit one end of the mounting rod 582 to prevent the mounting rod 582 from being separated from the step mounting hole 523. Specifically, one end of the mounting rod 582 can be connected to the second flange 584, so that the second flange 584 abuts against the first step surface 5231, thereby limiting the mounting rod 582. At the same time, one end of the second flange 584 away from the second step surface 5232 can abut against the adapter plate 524, so as to limit the extreme position of the mounting rod 582 through the adapter plate 524 and the first step surface 5231.

The pressing head 583 is connected to the other end of the mounting rod 582 and can extend out of the stepped mounting hole 523 to press the carrier 4 . Optionally, the mounting rod 582 is threadedly connected to the pressing head 583 .

The buffer member 581 is sleeved on the mounting rod 582, and one end of the buffer member 581 presses against the second step surface 5232 of the step mounting hole 523, and the other end presses against the pressing head 583, so that when the pressing head 583 presses against the carrier 4, the pressing head 583 elastically contacts the carrier 4, thereby preventing the pressing head 583 from damaging the carrier 4.

The structure of the extrusion mechanism 58 and the installation method of the extrusion mechanism 58 and the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 are not limited to this. For example, in other optional embodiments, a through hole can be opened on the base 5212, and the extrusion mechanism 58 is arranged in the through hole. The extrusion mechanism 58 can be connected to an extrusion drive member (not shown in the figure) such as a cylinder or an electric cylinder that drives the extrusion mechanism 58 to move. Optionally, the through hole extends along the first direction, and the extrusion drive member drives the extrusion mechanism 58 to reciprocate along the first direction. During use, before the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 clamp the carrier 4, the end of the extrusion mechanism 58 close to the carrier 4 is located in the through hole, so as to facilitate the placement of the carrier 5 on the nucleic acid amplification module 20; after the first temperature regulating mechanism 52 and the second temperature regulating mechanism 56 clamp the carrier 4, the extrusion drive member drives the extrusion mechanism 58 to extend out of the through hole, thereby extruding the extrusion cavity 46. The extrusion chamber 46 is squeezed by the extrusion mechanism 58, so that the gas and/or liquid (the liquid can be a reaction sample) in the extrusion chamber 46 applies pressure to the reaction sample in the amplification chamber 41, and then the reaction sample squeezes the corresponding cavity wall of the amplification chamber 41, so that the corresponding cavity wall of the amplification chamber 41 expands outward, so that the outer side of the cavity wall corresponding to the amplification chamber 41 is fully in contact with the nucleic acid amplification module 10. At the same time, the reaction sample is attached to the inner side of the cavity wall of the amplification chamber 41 under the pressure of the extrusion chamber 46, and because the outer side of the cavity wall of the amplification chamber 41 is in full contact with the nucleic acid amplification module 10, the amplification efficiency can be greatly improved.

As shown in Figures 22 and 23, the nucleic acid amplification module 10 further includes a positioning mechanism 57, which is connected to the first temperature adjustment mechanism 52 or the second temperature adjustment mechanism 56 and is used to position the carrier 4. Exemplarily, the positioning mechanism 57 is disposed on a carrier plate 561, and the positioning mechanism 57 is used to keep the side wall 42 of the carrier 4 at a preset distance from the nucleic acid detection module 20, thereby ensuring the accuracy of the fluorescence detection result.

The positioning mechanism 57 includes a first positioning component, which is used to position the carrier 4 in a direction perpendicular to the insertion direction of the first temperature adjustment mechanism 52 and the second temperature adjustment mechanism 56, as shown in Figure 22. In this embodiment, the first positioning component positions the carrier 4 in the first direction and the second direction, and the first direction and the second direction are perpendicular to the insertion direction of the carrier 4, and the insertion direction is consistent with the vertical direction.

Optionally, the first positioning assembly includes a first positioning portion 5711 and a second positioning portion 5712, and the first positioning portion 5711 and the second positioning portion 5712 are arranged at intervals along the second direction, so that the carrier 4 can be inserted between the first positioning portion 5711 and the second positioning portion 5712, and the carrier 4 can be restricted between the first positioning portion 5711 and the second positioning portion 5712. At the same time, when the carrier 4 is placed on the carrier plate 561, the position where the extrusion cavity 46 is provided on the carrier 4 is located between the first positioning portion 5711 and the second positioning portion 5712, and the extrusion mechanism 58 is also located between the first positioning portion 5711 and the second positioning portion 5712, that is, the first positioning portion 5711 and the second positioning portion 5712 will not block the extrusion mechanism 58 from pressing on the carrier 4, so that there will be no interference between the first positioning assembly and the extrusion mechanism 58.

Further, the first positioning portion 5711 and the second positioning portion 5712 are symmetrically arranged and both include a first plate 5713 and a second plate 5714. The first plate 5713 and the second plate 5714 are connected to form an L-shaped structure. One end of the first plate 5713 is connected to the carrier plate 561, and the other end is connected to the second plate 5714. One second plate 5714 extends in the direction of the other first plate 5713 and is arranged parallel to the carrier plate 561. The two first plates 5713 limit the carrier 4 in the second direction, and the two second plates 5714 and the carrier plate 561 limit the carrier 4 in the first direction.

The positioning mechanism 57 also includes a second positioning component 572 , which is disposed opposite to the nucleic acid detection module 20 . The second positioning component 572 and the nucleic acid detection module 20 are respectively abutted against two opposite side walls 42 of the carrier 4 .

Optionally, the second positioning assembly 572 includes a fixing portion 5721, an elastic adjustment portion 5722 and an abutting portion 5723. The fixing portion 5721 is connected to the carrier plate 561, one end of the elastic adjustment portion 5722 is connected to the fixing portion 5721, and the other end is connected to the abutting portion 5723, the abutting portion 5723 is used to abut the side wall 42 of the carrier 4, and the elastic adjustment portion 5722 can be a spring or the like. Optionally, the carrier 4 is inserted between the abutting portion 5723 and the optical fiber holder 2012 from the front end of the abutting portion 5723, and the front end of the abutting portion 5723 is provided with a chamfer so that the carrier 4 can smoothly enter between the abutting portion 5723 and the optical fiber holder 2012. When the carrier 4 is located between the abutting portion 5723 and the optical fiber holder 2012, the elastic adjustment portion 5722 is compressed to push the abutting portion 5723 to continuously abut against the carrier 4.

As shown in FIG. 22 , optionally, the thickness of the side of the carrier 4 close to the abutment portion 5723 is larger, so that the abutment portion 5723 can stably abut the carrier 4, and when the abutment portion 5723 abuts the carrier 4, deformation of the carrier 4 is avoided. To this end, the spacing between the second plate 5714 and the carrier plate 561 needs to allow the thicker side of the carrier 4 to pass through.

The thickness of the position where the extrusion cavity 46 is provided on the carrier 4 is greater than the thickness of the position where the amplification cavity 41 is provided, and is roughly the same as the thickness of the thicker side of the carrier 4. On the one hand, the strength of the carrier 4 at the position where it is squeezed by the extrusion mechanism 58 can be greater, thereby avoiding damage to the cavity wall corresponding to the extrusion cavity 46, and making the thickness of the extrusion cavity 46 larger, thereby having sufficient deformation space; on the other hand, the second plate 5714 and the supporting plate 561 can be used to position the carrier 4.

As shown in FIG. 18 and FIG. 24 a , in order to reduce the time required for cooling the heater 522 , optionally, the heater 522 is a thin layer structure, and the thickness of the heater 522 is approximately between a few tenths of a millimeter and several millimeters.

As shown in Fig. 24a, the heater 522 includes a heating element 91. A power source is connected to the heating element 91, which is a controllable heating source inside the heater 522 and can be a resistor, such as a copper material made into a resistor fine wire structure, and the heating power is controlled by controlling the current flowing through the resistor, thereby achieving temperature control. In other optional embodiments, the heating element 91 can also be a coil structure or electromagnetic induction heating through ferromagnetic materials, etc.

As shown in Figure 24a, preferably, heater 522 includes at least two independently controlled heating elements 91, and heating element 91 can be independently controlled to improve the uniformity of reaction sample temperature. As, if the temperature of a heating element 91 does not reach the preset temperature (how to detect the temperature of heating element 91 is described in detail below), the electric current of this heating element 91 is increased, and reaction sample is quickly raised to the preset temperature. In the present embodiment, because heater 522 is in direct contact with the reaction sample in carrier 4, the thickness of heater 522 is small, and the heat conduction efficiency between the reaction sample is high, the temperature of the heating element 91 of heater 522 can be equal to the temperature of reaction sample, therefore, the temperature of each heating element 91 is controlled to reach the preset temperature, reaction sample everywhere can be made to be in the preset temperature, and then the reaction sample temperature uniformity is guaranteed.

As shown in Fig. 24a, the heater 522 may further include an upper conductive component 92 and a lower conductive component 95, and the heating element 91 is sandwiched between the upper conductive component 92 and the lower conductive component 95. The upper conductive component 92 and the lower conductive component 95 have heat conduction and insulation functions.

The heater 522 includes a heat-scaling layer 921. Specifically, the upper conductive component 92 may also include a heat-scaling layer 921. The heat-scaling layer 921 contacts the first wall 42 or the second wall 43 of the carrier 4. The heat-scaling layer 921 can ensure uniform heat conduction in both the longitudinal and transverse directions (i.e., the thickness direction of the reaction sample and the surface perpendicular to the thickness direction), thereby ensuring the temperature uniformity of the sample liquid. Optionally, the upper conductive component 92 may also include an insulating layer 922. The insulating layer 922 may be made of insulating materials such as high thermal conductivity ceramics, and the heat-scaling layer 921 may be made of copper or aluminum.

The lower conducting component 95 also includes an insulating thermal resistance layer 951. The insulating thermal resistance layer 951 has certain thermal resistance characteristics and insulation characteristics. In addition to insulating the heating element 91, the insulating thermal resistance layer 951 can also form a longitudinal thermal resistance. The size of the thermal resistance can be designed by material selection and thickness selection to meet different design requirements. For example, a thin layer with a thickness of 0.1-0.3 mm can be used, and the thermal conductivity of the material is selected in the range of 0.2-0.5 W/mK.

Optionally, the lower conduction component 95 further includes a heat-conducting layer 952 made of copper or other heat-conducting materials, and the heat-conducting layer 952 is located on the side of the insulating thermal resistance layer 951 away from the heating element 91. Further, the heat-conducting layer 952 is the outermost layer of the lower conduction component 95, which is directly in contact with the cooling component 521. The heat-conducting layer 952 is made of metal such as copper or other materials with high thermal conductivity. Due to cost control or processing technology limitations, it is difficult to avoid point contact between the surface of the lower conduction component 95 and the cooling component 521. When the outermost layer of the lower conduction component 95 is the heat-conducting layer 952, even if there is point contact between the heat-conducting layer 952 and the cooling component 521, the heat-conducting layer 952 can also distribute the heat evenly throughout the heat-conducting layer 952 due to its good conductivity, thereby evenly distributing the heat of other layers of the lower conduction component 95.

Preferably, the heating element 91 of this embodiment is a resistance wire, and there is a specific relationship between the resistance wire and its temperature. Therefore, the real-time resistance change of the heating element 91 can be measured while heating, and the average temperature of the heating element 91 can be deduced through the resistance temperature coefficient and the nominal resistance value. The temperature reflects the current temperature of the heater 522 in real time without delay, so it can be used for rapid feedback control of the heater 522 and the reaction sample temperature. Compared with the prior art, the sample temperature can be controlled more accurately and the overall response speed of the temperature control system can be improved.

In order to detect the resistance of the heating element 91, the heater 522 may optionally further include a wiring port 96, which is used for a resistance detection element to detect the resistance of the heating element 91, so as to measure the temperature of the heating element 91 by a resistance temperature measurement method. The resistance detection element may include a horizontal patch connector (not shown in the figure), which has a plurality of connection terminals, which are inserted into the wiring port 96 and electrically connected to the wiring port 96, so that the voltage U and current I of the heating element 91 are detected through the wiring port 96, and the resistance R (R=U/I) is obtained. At the same time, the heating element 91 can also be powered by being electrically connected to the connection terminal through the wiring port 96.

Although the resistance temperature measurement method can measure the temperature of the carrier 4 without delay, the disadvantage of the resistance temperature measurement method is that for the same type of resistance wire (heating element 91), such as copper wire resistance wire, the nominal resistance value and resistance temperature coefficient (the resistance value at the nominal temperature is referred to as the nominal resistance value, the nominal resistance means that at this temperature, the declared (or marked) resistance value is real, where this temperature is the nominal temperature, and the nominal temperature can be selected arbitrarily according to demand) between the resistance wires are slightly different, resulting in the real resistance temperature coefficient of a single heating element 91 being slightly different from the nominal resistance value, which may cause temperature measurement errors. Therefore, preferably, as shown in Figure 24a, the heater 522 also includes a first temperature detection unit 99 for detecting the temperature of the heater 522. The first temperature detection unit 99 may include a contact temperature sensor or a non-contact temperature sensor, etc. The non-contact temperature sensor is an infrared sensor that does not need to be in contact with the heater 522 during temperature measurement. As shown in FIG. 24 b , the non-contact temperature sensor can be located in the through hole 5611 to facilitate detection of the temperature of the heater 522, and at the same time, the compactness of the structure of the nucleic acid amplification module 10 can be improved; the contact temperature sensor is a sensor that needs to be in contact with the heater 522 during temperature measurement. Although the first temperature detection unit 99 can detect the temperature of the heater 522, since the temperature of the reaction sample changes very quickly during the amplification stage, when the first temperature detection unit 99 detects the temperature of the heater 522, the first temperature detection unit 99 needs a certain reaction time to measure the temperature. Therefore, under normal circumstances, the detection result measured by the first temperature detection unit 99 will have a temperature measurement delay of 1 to 2 seconds. In the process of rapid temperature rise and fall, the temperature change of the heater 522 can reach more than 30°C in 1-2 seconds. Therefore, in the process of rapid temperature rise and fall, it is relatively difficult to control the heater 522 through the first temperature detection unit 99. In this embodiment, the heater 522 is controlled by a dual temperature measurement method of a resistance temperature measurement method and a first temperature detection unit 99 to calibrate the temperature of the heater 522 .

The present embodiment neither completely relies on the temperature value measured by the uncalibrated resistance temperature measurement method, nor completely relies on the temperature detected by the first temperature detection unit 99 to control the heater 522. Instead, the present embodiment combines the two, adopts the first temperature detection unit 99 to measure the temperature of the heater 522 and uses the resistance temperature measurement method to measure the temperature of the heating element 91, thereby being able to quickly and accurately control the temperature of the heater 522 and achieve the purpose of accurate temperature control, thereby overcoming the problems of temperature detection delay and large temperature measurement error caused by the temperature detection methods commonly used in the prior art.

In order to more clearly describe how to use the first temperature detection unit 99 to calibrate the temperature detected by the resistance detection element in this embodiment, in combination with FIG. 25, a process of calibrating the resistance temperature measurement method by the first temperature detection unit 99 in an actual detection is shown. Before calibrating the temperature value, an initial RT temperature curve, i.e., a temperature preset curve, is preset, and then a very small current is applied to the heating element 91 of the heater 522, such as a current of less than 1 mA. The purpose of applying a very small current is to read the resistance of the heating element 91 without causing the heating element 91 to heat up.

First calibration: the first temperature detection unit 99 measures the first temperature calibration value T, the resistance detection unit detects the first voltage _U1 and the first current _I1 of the heating element 91 at the temperature _T1 , and the resistance _R1 of the heating element 91 at the temperature _T1 can be obtained according to R=U/I.

Second calibration: The first temperature detection unit 99 then measures the second temperature calibration value T ₂ , and the resistance detection unit detects the second voltage U ₂ and the second current I ₂ of the heating element 91 at temperature T 2 , and the resistance R ₂ of the heating element 91 at temperature T ₂ can be obtained according to R=U/I.

Finally, according to two sets of two-variable linear equations: R ₁ =R ₀ (1+αΔT ₁ ) and R ₂ =R ₀ (1+αΔT ₂ ) (wherein, ΔT ₁ =T ₁ -T ₀ , ΔT ₂ =T ₂ -T ₀ , R ₁ is the resistance value of the heating element 91 corresponding to the temperature T ₁ , R ₂ is the resistance value of the heating element 91 corresponding to the temperature T ₂ , α is the resistance temperature coefficient of the material, T ₀ is the nominal temperature, and R ₀ is the nominal resistance value), the specific values of R0 and α are obtained, that is, the accurate RT curve is obtained, and the temperature of the heating element 91 measured by the resistance temperature measurement method can be used as feedback for accurate temperature control in the future.

The temperature calibration value can be detected throughout the entire process of nucleic acid amplification, so the temperature can be calibrated multiple times in the subsequent process to further improve the detection accuracy.

As shown in FIG. 24a , the heater 522 provided in this embodiment further includes a temperature calibration unit 93 for reflecting the temperature of the heating element 91, and the first temperature detection unit 99 detects the temperature of the temperature calibration unit 93. It can be understood that when multiple heating elements 91 are independently controlled, each heating element 91 is correspondingly provided with a temperature calibration unit 93, and each can be detected by a resistance detection unit to calibrate the heating element 91. The temperature calibration unit 93 can facilitate the measurement of the temperature of the carrier 4.

As shown in Figure 26, when the first temperature detection unit 99 is a contact temperature sensor, in order to facilitate the temperature measurement of the first temperature detection unit 99, the two first contacts of the first temperature detection unit 99 are respectively in contact with the two temperature calibration parts 93, and the two temperature calibration parts 93 are not conductive. At this time, optionally, the heater 522 can also include external electrical connection contacts 97 and electrical connection leads 98. The number of external electrical connection contacts 97 and electrical connection leads 98 can be two. The two external electrical connection contacts 97 are respectively located on the side away from each other of the two temperature calibration parts 93. One external connection contact is electrically connected to one temperature calibration part 93 through an electrical connection lead 98, and the other external connection contact is electrically connected to the other temperature calibration part 93 through another electrical connection lead 98.

As shown in Figure 24a, the heat-dissipating layer 921 of the upper conduction component 92 is conducted to the temperature calibration part 93, and the temperature calibration part 93 is electrically connected to the external resistance detection part at the external electrical connection contact 97 through the electrical connection lead 98. For example, the external resistance detection part can detect the resistance of the first temperature detection unit 99 through the external electrical connection contact 97, and then obtain the temperature of the first temperature detection unit 99 based on the resistance. Optionally, the diameter of the electrical connection lead 98 is smaller than that of the temperature calibration part 93 and the external electrical connection contact 97, thereby reducing the heat loss generated by the temperature calibration part 93 through the electrical connection lead 98, so that the temperature calibration part 93 can better reflect the temperature of the upper conduction component 92, such as the temperature of the heat-balancing layer 921 of the upper conduction component 92, and the first temperature detection unit 99 achieves good electrical and thermal contact with the temperature calibration part 93 through the solder joint. When the temperature of the upper conduction component 92, such as the heat-balancing layer 921 of the upper conduction component 92 changes, the first temperature detection unit 99 can quickly and accurately sense the temperature change. The temperature change causes the resistance value of the first temperature detection unit 99 to change. The resistance value change of the first temperature detection unit 99 is detected in real time at the external electrical connection contact 97, thereby achieving real-time temperature detection.

As shown in FIG. 20 , optionally, in order to shorten the time for the temperature of the temperature calibration part 93 to be consistent with the temperature of the heating element 91, the heater 522 may further include a fast conduction part 94, which is used to conduct the heat of the heating element 91 to the temperature calibration part 93. Specifically, in this embodiment, the heat of the heating element 91 is indirectly conducted to the temperature calibration part 93, such as the heating element 91 heats the heat-isolating layer 921, and the heat of the heat-isolating layer 921 is conducted to the temperature calibration part 93 through the fast conduction part 94, thereby, the temperature calibration part 93 accurately reflects the temperature of the heat-isolating layer 921, and then the first temperature detection unit 99 can accurately measure the temperature of the heat-isolating layer 921. Since the thickness of the reaction sample is very small, the temperature of the reaction sample is basically consistent with the temperature of the heat-isolating layer 921, so the temperature of the reaction sample can be obtained by detecting the temperature of the temperature calibration part 93.

Preferably, one side of the fast conduction part 94 is connected to one side of the upper conduction component 92 close to the heating element 91 or to one side of the lower conduction component 95 close to the heating element 91, and the other side is connected to the temperature calibration part 93. The lower surface of the upper conduction component 92 and the upper surface of the lower conduction component 95 are closest to the heating element 91, and their temperatures are the first to approach the temperature of the heating element 91. Therefore, the arrangement of the fast conduction part 94 can make the temperature of the fast conduction part 94 and the temperature of the heating element 91 reach the same level in the shortest time. Optionally, the fast conduction part 94 is made of a material with high thermal conductivity, such as a metal material such as copper or aluminum, or a thermally conductive ceramic. The thermal conductivity of the fast conduction part 94 is particularly better than that of the lower conduction component 95, so as to quickly transfer heat to the temperature calibration part 93.

The fast conduction part 94 includes a patch 941 and one or more guide pillars 942. The patch 941 is attached to the side of the upper conduction component 92 close to the heating element 91 or to the side of the lower conduction component 95 close to the heating element 91. One end of the one or more guide pillars 942 is connected to the patch 941, and the other end is passed through the lower conduction component 95 and connected to the temperature calibration part 93. The lower surface of the upper conduction component 92 and the upper surface of the lower conduction component 95 are closest to the heating element 91, and their temperatures are the first to approach the temperature of the heating element 91. Therefore, the arrangement of the patch 941 can make the temperature of the fast conduction part 94 consistent with the temperature of the heating element 91 as quickly as possible. The patch 941 can increase the contact area between the fast conduction part 94 and the upper conduction component 92 or the lower conduction component 95, thereby improving the conduction efficiency. The cross-sectional area of the guide pillar 942 can be smaller than the cross-sectional area of the patch 941, so that the temperature of the patch 941 can be quickly conducted to the temperature calibration part 93. Optionally, patch 941 and guide pillar 942 are made of materials with high thermal conductivity such as copper. When patch 941 and guide pillar 942 are required to be insulating materials to prevent heater 522 from short-circuiting, patch 941 or guide pillar 942 can be made of materials such as high thermal conductivity ceramics.

It is understandable that the temperature calibration part 93 can be arranged in one-to-one correspondence with the patch 941, and two temperature calibration parts 93 can also be connected to one patch 941. One temperature calibration part 93 can be connected to one guide post 942, and in order to improve the temperature uniformity of the temperature calibration part 93, the temperature calibration part 93 can also be connected to multiple guide posts 942.

In this embodiment, the wiring port 96 enables the heater 522 to realize its own temperature measurement function. Compared with the traditional structure that can only measure the temperature through an external temperature measurement unit, this embodiment can directly measure the temperature of the heater 522 itself, so the temperature measurement is more accurate and faster, which can improve the accuracy and control speed of the temperature control system.

The working process of the nucleic acid amplification module 10 provided in this embodiment is as follows: the transfer module 60 inserts the carrier 4 into the positioning mechanism 57, and the driving mechanism 53 drives the first temperature adjustment mechanism 52 to move to the side where the second temperature adjustment mechanism 56 is located, until the first temperature adjustment mechanism 52 and the second temperature adjustment mechanism 56 clamp the carrier 4, and at this time, the extrusion mechanism 58 presses to the position where the extrusion cavity 46 is provided on the carrier 4, thereby adjusting the pressure in the amplification cavity 41. The heater 522 and the cooling component 521 are opened and closed under the control of the control module 52 to achieve the amplification of the reaction sample in the carrier 4, the first temperature adjustment mechanism 52 moves in the direction away from the second temperature adjustment mechanism 56, and the transfer module 60 takes out the carrier 4. If the PCR automated integrated machine is used for the first time or after multiple uses, the heater 522 can also be temperature calibrated.

As shown in Fig. 2b and Fig. 27, optionally, at least part of the side wall 42 of the amplification chamber 41 is light-transmissive, and the nucleic acid detection module 20 detects the reaction sample through the light-transmissive side wall 42 of the carrier 4. Optionally, the material of the light-transmissive side wall 42 can be polydimethylsiloxane (i.e., PDMS), polypropylene (i.e., PP) or polycarbonate (i.e., PC), and PDMS, PP and PC are optically transparent materials with good biocompatibility.

Existing PCR automated all-in-one machines often choose a larger surface for nucleic acid detection, so that the reaction sample area receiving the excitation light is large, the generated fluorescence signal is strong, and accurate detection results are easy to obtain. According to the above thinking habits, due to the large area of the first wall 43 or the second wall 44 in this embodiment, fluorescence detection through the first wall 43 or the second wall 44 is often considered. However, in this embodiment, rapid temperature rise and fall can be achieved by setting a temperature adjustment mechanism on both sides of the carrier 4. The nucleic acid detection module 20 performs fluorescence detection on the side of the carrier 4. While achieving rapid temperature rise and fall and fluorescence detection, relative to the method of performing fluorescence detection through the first wall 43 or the second wall 44, the PCR automated all-in-one structure is more compact, reducing its large occupied space, and further improving the detection efficiency.

As shown in FIG28 , the fluorescence detection optical path in the prior art is an integrated structure, that is, the excitation light path 200 of the fluorescence detection optical path and the fluorescence signal optical path 201 are transmitted through an optical fiber, so that the excitation light emitted by the light source 205 enters the fluorescence signal optical path 201 after being reflected by the optical fiber. A dichroic mirror 203 and a filter 204 are arranged on the optical fiber transmission path, and both the dichroic mirror 203 and the filter 204 allow the fluorescence signal to pass through, and filter the excitation light in the fluorescence signal optical path 201. However, since the dichroic mirror 203 and the filter 204 do not filter the excitation light 100%, they also cannot allow the fluorescence signal to pass 100%, that is, the fluorescence signal will be attenuated when passing through the dichroic mirror 203 and the filter 204, and part of the filtered excitation light will enter the detection circuit 206 for detecting the fluorescence signal. In the prior art, since the area of the reaction sample excited by the excitation light is large and the number of generated fluorescent signals is also large, the excitation light passing through the filter 204 and the attenuated fluorescent signal have little effect on the detection result, and a more accurate detection result can be obtained.

However, the thickness of the amplification chamber 41 in this embodiment is thin (0.3-0.6 mm), and when the fluorescence detection is performed through the side wall 42 of the amplification chamber 41, the reaction samples that can receive the excitation light are greatly reduced, so the number of fluorescence signals excited each time is about one order of magnitude less. If the existing fluorescence detection optical path is used for detection, the signal detected by the detection circuit is low, and the excitation light passing through the filter will drown the fluorescence signal, which will lead to large errors in the detection, and even the fluorescence detection circuit cannot detect the fluorescence signal.

As shown in Figure 27, the nucleic acid detection module 20 provided in this embodiment includes a fluorescence detection optical path system 201, and the fluorescence detection optical path system 201 includes at least one fluorescence emission unit 1, and the fluorescence emission unit 1 is used to emit excitation light. Optionally, in this embodiment, at least two fluorescence emission units 1 are provided, and at least two fluorescence emission units 1 are used to emit excitation light respectively, and the control module 50 controls at least two fluorescence emission units 1 to emit excitation light in different time periods respectively. Of course, the number of fluorescence emission units 1 can also be one. Exemplarily, as shown in Figure 27, four fluorescence emission units 1 can be provided. Of course, the number of fluorescence emission units 1 is not limited to four, and can also be less than four or more than four.

Optionally, the control module 50 is arranged on the frame 40. The control module 50 is also electrically connected to the nucleic acid amplification module 10 and the nucleic acid detection module 20 to control the nucleic acid amplification module 10 and the nucleic acid detection module 20. The control module 50 can also be electrically connected to each other module to control each module separately. It is understandable that the control module 50 can be a centralized or distributed control module 50. For example, the control module 50 can be a single microcontroller or a distributed multi-chip microcontroller. The control program can be run in the microcontroller to control each component to realize its function. The control module 50 can be connected to each module through a line or through a wireless connection such as Bluetooth.

The fluorescence emission unit 1 is independently arranged, and the excitation light transmitted in the fluorescence emission unit 1 will not be reflected into the optical path of the fluorescence signal, thereby greatly reducing the amount of excitation light in the optical path of the fluorescence signal, greatly reducing the background in the fluorescence signal, and improving the detection accuracy. At the same time, when the reaction sample in the carrier 4 has a variety of fluorescent probes/dyes, the controller can control the opening of different fluorescence emission units 1 in sequence in different time periods, such as the first fluorescence emission unit 1 works within the 1st-2s from the start of the detection, and the second fluorescence emission unit 1 works within the 2nd-3s from the start of the detection, and so on. Only one fluorescence emission unit 1 works in a time period, and a fluorescence emission unit 1 only needs to emit one excitation light. Therefore, only the type of excitation light is single in a time period, and the type of fluorescent signal is single. When detecting the fluorescent signal, the noise generated by the excitation light is small, which can further improve the accuracy of the detection result. In addition, by controlling the conduction of the fluorescence emission unit 1 of each channel 49 by the controller, only one excitation light is achieved at the same time, and the switching of the excitation light channel 49 is achieved in milliseconds, which can realize rapid detection of the reaction sample.

Optionally, the wavelengths of the excitation lights emitted by at least two fluorescence emission units 1 are different from each other to improve the utilization rate of the fluorescence emission units 1. Of course, in order to avoid inaccurate detection results caused by failure of the fluorescence emission unit 1, a spare fluorescence emission unit 1 can also be provided, such as in at least two fluorescence emission units 1, the wavelengths of the excitation lights emitted by the two fluorescence emission units 1 are the same.

As shown in Figure 27, the fluorescence emission unit 1 includes a light source 12 and an excitation optical fiber 11. The light source 12 can be an LED, etc. The light source 12 is used to emit excitation light, and the light source 12 is electrically connected to the control module 50. The excitation optical fiber 11 is used to transmit the excitation light emitted by the light source 12 to transmit the excitation light to the reaction sample. The light source 12 and the reaction sample are connected by the excitation optical fiber 11. The excitation optical fiber 11 is convenient for isolating the heat source, making the experimental data more stable, and the connection between the excitation optical fiber 11 and the light source 12 is simple, which is conducive to earthquake resistance. At the same time, since the optical fiber itself is small in size, it can meet the needs of the flat structure carrier 4. In addition, one light source 12 corresponds to one excitation optical fiber 11, and each excitation channel 49 has a separate optical fiber, which is easy to couple and has small light loss.

As shown in Fig. 27, the fluorescence detection optical path system 201 also includes a fluorescence detection unit 2. The fluorescence detection unit 2 is used to detect the fluorescence signal, and the fluorescence emission unit 1 is completely separated from the fluorescence detection unit 2 that receives the excitation signal, thereby reducing the background of the fluorescence signal and improving the detection sensitivity of the fluorescence detection optical path system 201. At the same time, the fluorescence emission unit 1 is completely separated from the fluorescence detection unit 2, reducing the optical elements in the fluorescence detection unit 2, such as the dichroic mirror, etc., thereby reducing the cost and improving the detection efficiency of the fluorescence.

As shown in Figure 29, when the number of fluorescence detection units 2 is one, the fluorescence detection unit 2 includes a detector 22, a fluorescence transmission optical path 21 and a turntable 23. A plurality of filters 211 are arranged on the turntable 23. The fluorescence transmission optical path 21 includes an optical fiber 212. A plurality of fluorescence signals are transmitted to the filter 211 through the optical fiber 212. The plurality of filters 211 can pass a fluorescence signal respectively. The channels are switched by rotating the turntable 23 so that different fluorescence signals can pass through and then be received by the detector 22.

At the same time, the present embodiment provides a more preferred solution of the fluorescence detection unit 2, as shown in Figures 27 and 30, the fluorescence detection unit 2 includes at least two fluorescence transmission optical paths 21, and the at least two fluorescence transmission optical paths 21 correspond one-to-one to the at least two fluorescence emission units 1, that is, one fluorescence emission unit 1 can correspond to one fluorescence transmission optical path 21. The reaction sample at the location opposite to the excitation optical fiber 11 of the fluorescence emission unit 1 can receive stronger excitation light, so the reaction sample at this location can generate a higher fluorescence signal. The at least two fluorescence transmission optical paths 21 correspond one-to-one to the at least two fluorescence emission units 1, so that the fluorescence transmission optical path 21 can be set close to the reaction sample excited by the corresponding fluorescence emission unit 1, thereby allowing more fluorescence signals to enter the fluorescence transmission optical path 21, further improving the accuracy of the detection result.

Each fluorescence transmission optical path 21 includes a filter 211 that allows a preset fluorescence signal to pass through, that is, each fluorescence transmission optical path 21 can only allow a specific fluorescence signal to pass through. Compared with the above-mentioned setting of only one fluorescence transmission optical path 21, by mechanically switching different filters 211, a filter 211 is set in each fluorescence transmission optical path 21, which can reduce the time required for mechanical switching when the fluorescence signal passes through the optical path, thereby realizing rapid detection of the fluorescence signal and eliminating the influence of mechanical vibration caused by mechanical switching on the detection result.

As shown in Figures 27 and 30, the fluorescence detection unit 2 also includes a detector 22, and the multiple fluorescence transmission optical paths 21 are all connected to the detector 22, and the detector 22 is used to detect the fluorescence signal. The detector 22 includes a silicon photomultiplier tube (i.e., SiPM), a photon type detector (i.e., PD) or a photomultiplier tube (i.e., PMT). The fluorescence detector 22 using the silicon photomultiplier tube, the photon type detector or the photomultiplier tube has high sensitivity. When performing fluorescence detection, ultra-fast and high-sensitivity detection of the fluorescence signal at the millisecond level is achieved.

Preferably, the number of detectors 22 is one, and the detector 22 is electrically connected to the control module 50 to record the intensity of the fluorescence signal in different time periods. Different fluorescence transmission optical paths 21 converge on one detector 22 after passing through different filters 211. The fluorescence emission unit 1 turns on different fluorescence emission units 1 in different time periods to switch the excitation channel 49, thereby exciting different signal fluorescence in different time periods. The corresponding signal fluorescence can pass through the corresponding filter 211. Although the fluorescence detector 22 can only detect the intensity of the signal fluorescence but not the type of the signal fluorescence, it can determine what kind of signal fluorescence it is according to the receiving time period, and then determine the reaction sample corresponding to the signal fluorescence. Compared with the scheme of Figure 29, the above technical scheme does not require a motor or the like to drive the turntable 23 to rotate, and there is no mechanical switching, which avoids vibration caused by the rotation of the turntable 23, and improves the accuracy of the detection result. According to the time period, it is determined what kind of signal fluorescence it is, the switching between fluorescence channels is fast, the detection speed is fast, the detection efficiency is high, the equipment cost is low, and the equipment structure is simple.

As shown in FIG. 27 and in combination with FIG. 23 , the fluorescence transmission optical path 21 includes a collection optical fiber 212 , an emission end of the excitation optical fiber 11 for emitting excitation light, and an injection end of the collection optical fiber 212 for injecting fluorescence signals to form an optical fiber group 2011 .

As shown in Fig. 31 and Fig. 32, and in combination with Fig. 23, the fluorescence detection optical path system 100 further includes a fiber seat 2012, and the fiber group 2011 is disposed in the fiber seat 2012. As shown in Fig. 31 and Fig. 32, the fiber seat 2012 includes a seat body 2013 and a flat groove 2014 opened on one side of the seat body 2013, and the fiber group 2012 is disposed in the flat groove 2014. In combination with Fig. 23, the second positioning assembly 572 is disposed opposite to the fiber seat 2012, and the second positioning assembly 572 and the fiber seat 2012 are respectively abutted against the opposite side walls 42 of the carrier 4.

The optical fibers in the optical fiber group 2011 are arranged along a preset direction, which is the radial direction of the optical fibers (the direction indicated by the arrow R in FIG. 27 is the radial direction of the optical fibers). In this embodiment, the preset direction is also the vertical direction (as shown in FIG. 23 ), so that the emission end of the collection optical fiber 212 and the injection end of the excitation optical fiber 11 are arranged flat in the optical fiber holder 2012, as shown in FIG. 23 , that is, the optical fibers (collection optical fiber 212 and excitation optical fiber 11) are arranged flat at one end close to the side wall 42 of the carrier 4. Exemplarily, the preset direction is the length direction of the side wall 42. In the thickness direction of the carrier 4, the optical fibers in the optical fiber holder 2012 are arranged in a single layer, so as to adapt to the flat structure of the carrier 4. It can be understood that the diameter of the optical fiber can be smaller than the thickness of the carrier 4, so that the optical fiber in the optical fiber holder 2012 does not protrude from the carrier 4 in the thickness direction of the carrier 4, so that the excitation light emitted by the collection optical fiber 212 can enter the accommodating cavity 41, and the entire excitation optical fiber 11 can collect fluorescence. The excitation optical fiber 11 and the collection optical fiber 212 are arranged in parallel to obtain the best sensitivity and signal-to-noise ratio. At the same time, when the optical fiber group 2011 is coupled to the carrier 4, the preset direction of the optical fiber arrangement is perpendicular to the thickness direction of the carrier 4, and the optical fiber group 2011 and the optical fiber holder 2012 are both flat structures, so as to match the carrier 4.

As shown in Figure 33, when at least two fluorescence transmission optical paths 21 and at least two fluorescence emission units 1 are provided, at least two groups of optical fiber groups 2011 are formed accordingly, and at least two groups of optical fiber groups 2011 in the optical fiber holder 2012 are arranged in sequence along a preset direction, that is, when multiple optical fiber groups 2011 are provided, the multiple optical fiber groups 2011 are also arranged in a flat manner, and in the thickness direction of the carrier 4, the optical fibers in the optical fiber holder 2012 are also arranged in a single layer, so as to adapt to the flat structure of the carrier 4, and the optical fibers in the optical fiber holder 2012 do not protrude from the carrier 4 in the thickness direction of the carrier 4, therefore, the excitation light emitted by the collecting optical fiber 212 can enter the accommodating cavity 41, and the entire excitation optical fiber 11 can collect fluorescence.

At the same time, multiple optical fiber groups 2011 are arranged in sequence, and the collection optical fiber 212 and the excitation optical fiber 11 in the same optical fiber group 2011 are adjacent to each other. The reaction sample at the corresponding position of the excitation optical fiber 11 can generate more fluorescence signals, and the collection optical fiber 212 is close to the corresponding excitation optical fiber 11, so that more fluorescence signals can enter the fluorescence transmission optical path 21, thereby further improving the accuracy of the detection results.

As shown in FIG. 33 , a fiber group 2011 includes at least two collecting fibers 212, and at least one collecting fiber 212 is arranged on both sides of the preset direction of the excitation fiber 11 in the fiber group 2011. In a fiber group 2011, increasing the number of collecting fibers 212 to at least two and performing multi-point detection on the reaction sample can effectively improve the detection efficiency of fluorescence, solve the problem of low signal, and reduce the demand for the sensitivity of the detector 22. At the same time, when one of the collecting fibers 212 is affected by bubbles in the reaction sample, it can be corrected by the detection results of other collecting fibers 212. Exemplarily, a collecting fiber 212 is arranged on both sides of an excitation fiber 11, and a fiber group 2011 is formed, that is, each fluorescence can be collected by two collecting fibers 212. Of course, only one collecting fiber 212 or more than two collecting fibers 212 can also be arranged in a fiber group 2011.

As shown in FIG33 , a fluorescence emission unit 1 may be provided only on one side of the carrier 4, or, as shown in FIG34 , a fluorescence emission unit 1 may be provided on both sides of the carrier 4, and a collection fiber 212 may be provided corresponding to the excitation fiber 11 of a fluorescence emission unit 1, and each collection fiber 212 on both sides of the carrier 4 is preferably connected to a detector 22, and of course, the collection fiber 212 on each side may be connected to a detector 2222 respectively. Detection on both sides of the reaction sample can effectively improve the detection efficiency of fluorescence, solve the problem of low signal, and reduce the demand for the sensitivity of the detector 22. Moreover, when the detection result of the collection fiber 212 on one side is inaccurate due to the obstruction of bubbles in the reaction sample, etc., it can be corrected by the detection result on the other side. Of course, detection can also be performed on three sides or more than three sides of the carrier 4, and preferably, fluorescence detection is performed through the light-transmitting side wall 42 of the carrier 4.

Exemplarily, when the carrier 4 and the reagent carrying part 1001 are an integrated structure, and the thickness direction of the carrier 4 is consistent with the vertical direction, the working process of the PCR automated integrated machine provided in this embodiment is:

Manually or automatically placing the reagent kit 100 on the bracket 311;

The control module 50 controls the pipetting submodule 32 to configure the sample and reagents in the reagent kit 100 into a reaction sample;

The cutting module 70 cuts the reagent kit 100 to separate the carrier 4 and the reagent carrying portion 1001 , and the flipping module 7 flips the carrier 4 90°; or the bending module 80 bends and breaks the reagent kit 100 ;

The control module 50 controls the transfer module 60 to transfer the carrier 4 to the nucleic acid amplification module 10, and the nucleic acid amplification module 10 works to achieve amplification of the reaction sample;

The control module 50 controls the nucleic acid detection module 20 to detect the reaction sample;

The control module 50 controls the transfer module 60 to transfer the reagent carrying part 1001 to the recovery module 90;

The control module 50 controls the transfer module 60 to take the carrier 4 out of the nucleic acid amplification module 10 and transfer the carrier 4 to the recovery module 90 .

Exemplarily, when the carrier 4 and the reagent carrying part 1001 are an integrated structure, and the thickness direction of the carrier 4 is consistent with the vertical direction, the working process of the PCR automated integrated machine provided in this embodiment can also be:

Manually or automatically placing the reagent kit 100 on the bracket 311;

The control module 50 controls the pipetting submodule 32 to configure the sample and reagents in the reagent kit 100 into a reaction sample;

The bending module 80 bends the carrier 4 90°, but the reagent box 100 is not broken;

The control module 50 controls the transfer module 60 to transfer the reagent kit 100 to the nucleic acid amplification module 10, and the nucleic acid amplification module 10 works to achieve amplification of the reaction sample;

The control module 50 controls the nucleic acid detection module 20 to detect the reaction sample;

The control module 50 controls the transfer module 60 to take the carrier 4 out of the nucleic acid amplification module 10 and transfer the carrier 4 to the recovery module 90 .

Exemplarily, when the carrier 4 and the reagent carrying part 1001 are separate structures, the working process of the PCR automatic integrated machine provided in this embodiment is as follows:

Manually or automatically placing the reagent kit 100 on the bracket 311;

The control module 50 controls the pipetting submodule 32 to configure the sample and reagents in the reagent kit 100 into a reaction sample;

The flip module 7 flips the carrier 4 90° (when the thickness direction of the carrier 4 is consistent with the vertical direction, this step is included; when the thickness direction of the carrier 4 is perpendicular to the vertical direction, this step is not included);

The control module 50 controls the transfer module 60 to transfer the carrier 4 to the nucleic acid amplification module 10, and the nucleic acid amplification module 10 works to achieve amplification of the reaction sample;

The control module 50 controls the nucleic acid detection module 20 to detect the reaction sample;

The control module 50 controls the transfer module 60 to transfer the reagent carrying part 1001 to the recovery module 90;

The control module 50 controls the transfer module 60 to take the carrier 4 out of the nucleic acid amplification module 10 and transfer the carrier 4 to the recovery module 90 .

Although the present invention has been described in detail above by general description, specific implementation methods and experiments, it is obvious to those skilled in the art that some modifications or improvements can be made on the basis of the present invention. Therefore, these modifications or improvements made on the basis of not departing from the spirit of the present invention all belong to the scope of protection claimed by the present invention.

Claims (35)

1. The automatic PCR all-in-one machine is characterized by comprising a rack, and a nucleic acid amplification module and a nucleic acid detection module which are arranged on the rack, wherein the nucleic acid amplification module is used for amplifying a reaction sample, and the nucleic acid detection module is used for detecting the reaction sample.

2. The automated PCR machine of claim 1, further comprising a compounding module disposed in the frame for compounding the sample with the reagent into a reaction sample.

3. The automated PCR integration machine of claim 2, wherein the preparation module includes a carrier sub-module and a pipetting sub-module disposed on an upper side of the carrier sub-module, the carrier sub-module and the pipetting sub-module being capable of moving relative to each other to allow the pipetting sub-module to transfer the solution on the carrier sub-module and to inject the reaction sample into the kit.

4. The PCR automation all-in-one of claim 3, the pipetting submodule including a pipetting gun, at least one of the pipetting gun and the carrier submodule being movable in a horizontal plane, at least one of the pipetting gun and the carrier submodule being movable in a vertical direction.

5. The automated PCR integration machine of claim 3, wherein the carrier sub-module is capable of carrying a kit and the pipetting sub-module is configured to transfer solutions between the chambers of the kit.

6. The automated PCR machine of claim 3, wherein the carrier sub-module includes a bracket having a receptacle for receiving a kit.

7. The automated PCR machine of claim 6, wherein the receptacle includes a plurality of receptacles into which the reagent carrying portion of the kit can be inserted.

8. The automated PCR machine of claim 7, wherein a heating structure is disposed within at least one of the plurality of receptacles for heating the reaction sample within the reagent carrier.

9. The automated PCR integration machine of claim 3, further comprising a transfer module disposed on the rack for transferring a kit or a vector between the carrier sub-module and the nucleic acid amplification module or between the carrier sub-module, the nucleic acid amplification module and the nucleic acid detection module.

10. The automated PCR machine of claim 1, further comprising a cutting module disposed in the frame for cutting off the kit between the carrier of the kit and the reagent carrying portion of the kit.

11. The automated PCR machine of claim 1 or 10, further comprising a flipping module disposed on the frame for flipping the carriers of the kit a predetermined angle.

12. The automated PCR machine of claim 1, further comprising a bending module disposed on the frame, the bending module configured to bend one end of the kit by a predetermined angle, or the bending module configured to bend a carrier of the kit by a predetermined angle and break the kit between the carrier and the reagent carrying portion thereof.

13. The automated PCR integration machine of claim 1, wherein the reaction sample is contained in a carrier, the carrier being of a flat configuration, the nucleic acid amplification module and/or the nucleic acid detection module having the carrier inserted vertically.

14. The automated PCR integration machine of claim 1, further comprising a control module for controlling the nucleic acid amplification module and the nucleic acid detection module.

15. The PCR automation all-in-one of claim 14, the nucleic acid detection module comprising a fluorescence detection light path system comprising:

at least one fluorescent emission unit for emitting excitation light.

16. The automated PCR machine of claim 15, wherein the number of fluorescent light emitting units is at least two, and the control module controls the at least two fluorescent light emitting units to emit excitation light at different time periods, respectively.

17. The automated PCR machine of claim 16, wherein the fluorescence detection light path system further comprises:

the fluorescent detection unit comprises at least two fluorescent transmission light paths, and at least two fluorescent emission units are in one-to-one correspondence with at least two fluorescent transmission light paths.

18. The automated PCR machine of claim 17, wherein the fluorescence detection unit further comprises a detector, and wherein the plurality of fluorescence transmission light paths are each coupled to the detector.

19. The automated PCR machine of claim 18, wherein the number of detectors is one, the detectors being electrically connected to the control module to record the intensity of the fluorescent signal over time.

20. The automated PCR machine of claim 17, wherein the fluorescence detection light path system further comprises a fiber holder, the fluorescence transmission light path comprises a collection fiber, the excitation fiber emits the emission end of the excitation light, and the emission end of the collection fiber emits the fluorescence signal forms a fiber group, the fiber group is disposed in the fiber holder, and the fibers in the fiber holder are arranged along a preset direction, and the preset direction is a radial direction of the fibers, such that the emission end of the collection fiber and the emission end of the excitation fiber are arranged flat in the fiber holder.

21. The automated PCR machine of claim 20, wherein at least the two fluorescence transmission light paths and the at least two fluorescence emission units form at least two sets of the optical fibers, the at least two sets of the optical fibers being arranged in sequence along the predetermined direction.

22. The automated PCR machine of claim 21, wherein one of the fiber sets includes at least two of the collection fibers, at least one of the collection fibers being disposed on either side of the predetermined direction of the excitation fibers in one of the fiber sets.

23. The automated PCR integration machine of any one of claims 1-10, 12-20, wherein the nucleic acid amplification module includes at least one set of temperature adjustment mechanisms that are capable of changing the temperature of the reaction sample within the carrier.

24. The automated PCR integration machine of claim 23, wherein the nucleic acid amplification module includes two sets of temperature adjustment mechanisms, a first temperature adjustment mechanism and a second temperature adjustment mechanism, respectively, the first temperature adjustment mechanism and/or the second temperature adjustment mechanism being adjustable in position to be closer to or farther from each other, and a carrier being sandwiched between the first temperature adjustment mechanism and the second temperature adjustment mechanism.

25. The automated PCR machine of claim 24, wherein the nucleic acid amplification module includes a drive mechanism that drives the first temperature adjustment mechanism and/or the second temperature adjustment mechanism toward or away from each other.

26. The automated PCR machine of claim 25, wherein a plurality of sets of resilient leveling assemblies are disposed between the drive mechanism and the first and/or second temperature adjustment mechanisms.

27. The automated PCR integration machine of claim 24, wherein the nucleic acid amplification module further includes a squeeze mechanism, the first and/or second temperature adjustment mechanisms being coupled to the squeeze mechanism to squeeze a squeeze cavity of a carrier sandwiched between the first and second temperature adjustment mechanisms.

28. The automated PCR machine of claim 27, wherein the first temperature adjustment mechanism and/or the second temperature adjustment mechanism has a step mounting hole formed therein, the extrusion mechanism includes an extrusion assembly and a buffer, the extrusion assembly is connected to the step mounting hole, and the buffer is connected between the extrusion assembly and the step mounting hole, so that the extrusion assembly is in elastic contact with a carrier.

29. The PCR automation all-in-one of claim 24, the first temperature adjustment mechanism and the second temperature adjustment mechanism each include a cooling assembly for cooling the carrier.

30. The PCR automation all-in-one machine of claim 29, the first and second temperature adjustment mechanisms each further comprising a heater disposed on a side of the cooling assemblies of the first and second temperature adjustment mechanisms that are proximate to each other.

31. The automated PCR machine of claim 30, wherein the heater comprises a heating element, and wherein the automated PCR machine further comprises a resistance detection element for detecting a resistance of the heating element.

32. The automated PCR machine of claim 31, further comprising a first temperature detection unit for detecting the heater temperature.

33. The automated PCR machine of claim 24, wherein the nucleic acid amplification module further comprises a positioning mechanism for positioning a carrier, the positioning mechanism coupled to the first temperature adjustment mechanism or the second temperature adjustment mechanism.

34. The automated PCR machine of claim 13, wherein the flat structure means that the ratio of the dimension of the carrier perpendicular to the thickness direction thereof to the dimension of the carrier perpendicular to the thickness direction thereof is greater than 5:1.

35. The automated PCR machine of claim 34, wherein the ratio of dimensions is 50:1 to 100:1.