CN110331092A - Nucleic acid total analysis micro-fluidic chip system and use method thereof - Google Patents

Abstract

The invention discloses a nucleic acid total analysis micro-fluidic chip system and a using method thereof, belonging to the technical field of micro-fluidic chip nucleic acid analysis. The nucleic acid full-analysis microfluidic chip comprises a nucleic acid full-analysis microfluidic chip and a portable device, wherein the microfluidic chip comprises: the nucleic acid analysis device comprises a plurality of nucleic acid analysis channels which are arranged in parallel, wherein each channel comprises a three-dimensional mixing pool and a plurality of reaction pools, and an isolation pool is arranged between every two adjacent reaction pools; the portable device comprises: the device comprises a rotating three-dimensional magnetic field control module, a one-dimensional moving magnetic field control module, a temperature control module and a signal detection module. The nucleic acid total analysis micro-fluidic chip and the portable device can realize the high-efficiency mixing and reaction process of multi-step magnetic beads, are easy to integrate other functional units, complete the automatic, low-cost and high-sensitivity nucleic acid micro total analysis process, and have wide application prospect.

Description

A microfluidic chip system for full analysis of nucleic acid and its application method

technical field

The invention relates to a microfluidic chip system for full nucleic acid analysis and a method for using the same, belonging to the technical field of microfluidic chip nucleic acid analysis.

Background technique

Nucleic acid analysis plays an important role in biology, medicine, pharmaceutical research, and disease prevention, diagnosis, and treatment. Most conventional nucleic acid analysis methods are based on PCR amplification of target fragments. The whole process also includes multi-step operations such as sample processing before amplification (such as cell lysis, nucleic acid purification and extraction), detection and analysis of amplified products, etc. One set of instruments and equipment is required, and professional operation is required. Various sample processing processes and product detection processes are often carried out separately, which is difficult to achieve automation and low consumption, resulting in low analysis efficiency and detection accuracy, and is not suitable for on-site rapid detection requirements. The unit technologies required for nucleic acid analysis can be transferred to the microfluidic chip in a single or integrated manner, and finally the basic operating units involved in a complete nucleic acid analysis process are integrated into the same microfluidic chip system to form a fully integrated The microfluidic chip nucleic acid analysis system will completely change the characteristics of time-consuming, laborious, poor automation, and low accuracy of traditional nucleic acid analysis methods. It can effectively reduce sample loss and pollution, significantly reduce analysis time and cost, and complete rapid and automated analysis. , has important theoretical and practical significance. The research on the microfluidic chip system for the full analysis of nucleic acid has been reported at home and abroad, but so far, most of the existing methods have single functions, are "tiny and incomplete", and have not been automated, or although the integration level is high, the chips and devices The structure is complex, the cost is high, the efficiency is low, and the throughput is insufficient, resulting in low precision, poor repeatability and poor practicability of the test results.

Contents of the invention

In order to solve the above problems, the present invention provides a nucleic acid full analysis microfluidic chip system and its use method, and specifically provides a nucleic acid full analysis microfluidic chip based on the magnetic bead method, a portable device and its use method. The method is especially suitable for automatic rapid detection of nucleic acid samples, and can be applied in special environments.

In order to achieve the above object, the present invention provides a microfluidic chip system for full nucleic acid analysis, including a rotating three-dimensional magnetic field control module 1, a one-dimensional moving magnetic field control module 2, a temperature control module 3, a signal detection module 4 and a microfluidic chip 5, of which:

The three-dimensional rotating magnetic field control module 1 includes an upper disc 11, an upper disc magnet 12, a lower disc 13, a lower disc magnet 14, a drive motor 15, a rotating shaft 16 and a rotating shaft base 17; the upper disc 11 and The lower circular plate 13 is arranged parallel to the corresponding level, and the upper circular plate 11 and the lower circular plate 13 are provided with a rotating shaft 16, and the rotating shaft 16 is arranged at a position deviated from the center of the circle and vertically runs through the upper circular plate 11 and the lower circular plate. 13; the top of the rotating shaft 16 is provided with a driving motor 15, and the bottom of the rotating shaft 16 is provided with a rotating shaft base 17, and the upper circular plate 11 and the lower circular plate 13 are driven by the driving motor 15 through the rotating shaft 16; the upper circular plate 11 and the edge of the lower disc 13 are respectively provided with the same shape, evenly arranged a plurality of upper disc magnets 12 and lower disc magnets 14, the long sides of the upper disc magnets 12 and the lower disc magnets 14 are in line with the The rotating shaft 16 is vertical, and the upper disc magnet 12 and the lower disc magnet 14 are staggered in the vertical position;

The one-dimensional mobile magnetic field control module 2 includes a microfluidic chip 5, a rectangular permanent magnet fixing assembly 21 and a microfluidic chip mobile control assembly 22; one end of the microfluidic chip 5 is provided with a three-dimensional mixing pool 51, the The three-dimensional mixing pool 51 is located in the magnetic field area between the upper circular plate 11 and the lower circular plate 13, and the side of the other end of the microfluidic chip 5 is provided with a microfluidic chip movement control assembly 22, and the microfluidic chip The lower end of the chip 5 is provided with a rectangular permanent magnet fixing component 21 adjacent to one side of the microfluidic chip mobile control component 22, and the other side of the rectangular permanent magnet fixing component 21 is provided with a temperature control module 3, and the temperature control module 3 Close to the lower end of the microfluidic chip 5 , the upper end of the microfluidic chip 5 is provided with a signal detection module 4 , and the signal detection module 4 is located above the temperature control module 3 .

Further, in the above technical solution, there are at least two channels on the microfluidic chip 5, the channels are arranged in parallel, and each channel is provided with a three-dimensional mixing pool 51 and at least two reaction pools in turn. 52, an isolation pool 54 is provided between the three-dimensional mixing pool 51 and the reaction pool 52, an isolation pool 54 is provided between the adjacent reaction pools 52, and the three-dimensional mixing pool 51, the reaction pool 52 and the isolation pool 54 Liquid feeding holes 53 are provided on each of them. One end point of the channel is a three-dimensional mixing pool 51 , and the other end point of the channel is an isolation pool 54 .

Further, in the above technical solution, the diameters of the upper circular plate 1 and the lower circular plate 3 are both 45 mm and the thickness is 3 mm, the eccentricity of the rotating shaft 6 is 7.5 mm, and the upper circular plate magnet 2 and the lower circular plate The disc magnet 4 is an NdFeB permanent magnet, and the upper disc magnet 2 and the lower disc magnet 4 are 8.5mm long, 1.5mm wide and 5mm high.

Further, in the above technical solution, the three-dimensional mixing pool 51 is a closed structure with a depth > 1 mm; the reaction pool 52 is flat and has a depth < 0.5 mm, and the volume ratio of the three-dimensional mixing pool 51 to the reaction pool 52 > 4:1.

Further, in the above technical solution, the temperature control area of the temperature control module 3 covers the reaction pool 52, and the temperature control module 3 includes a temperature control element and a control circuit.

Further, in the above technical solution, the detection area of the signal detection module 4 covers the reaction pool 52, and the signal detection module 4 includes a signal acquisition unit and a data processing unit.

The present invention also provides a method for using a microfluidic chip nucleic acid full analysis system, which includes the following steps:

① Add surface-modified magnetic beads and nucleic acid samples to be detected into the three-dimensional mixing pool 51, add corresponding reagents into the reaction pool 52, add spacer medium into the isolation pool 54, and then seal the filling holes of each liquid pool with a sealing film;

② Place the three-dimensional mixing pool 51 completely in the rotating three-dimensional magnetic field control area, start the rotating three-dimensional magnetic field control module 1, and drive the upper circular plate 11 and the lower circular plate 13 to rotate by driving the rotating shaft 16, so that the magnetic beads and the nucleic acid sample are fully contacted and mixed;

③ After step ② is completed, turn off the rotating three-dimensional magnetic field control module 1, start the microfluidic chip movement control component 22, and make the magnetic beads leave the three-dimensional mixing pool and enter the next reaction pool through the isolation pool;

④ After step ③ is completed, control the microfluidic chip to move the control component 22, so that the magnetic beads perform one-dimensional reciprocating motion in the reaction tank, so as to realize the sufficient mixing and reaction of the magnetic beads and the reagents in the reaction tank;

⑤After step ④ is completed, control the mobile control component 22 of the microfluidic chip, so that the magnetic beads enter the subsequent reaction pools in sequence, complete the subsequent mixing and reaction process in sequence, and perform reaction and detection in the final reaction pool, so as to realize nucleic acid completeness. Analysis process.

Further, in the above technical solution, the surface-modified magnetic beads in step ① include general-purpose magnetic silica beads, specific DNA probe-modified magnetic beads, immunomagnetic beads, and the like.

Further, in the above technical solution, the reagents added to the reaction pool in step ① include washing buffer, elution buffer, nucleic acid amplification reagents, and detection reagents.

Further, in the above technical solution, the isolation medium in step ① is another flow medium that is incompatible with water, and the other flow medium that is incompatible with water includes air, mineral oil, etc., to realize adjacent Separation of different aqueous solutions in the reaction tank.

The present invention has the following beneficial effects:

1. For the first time, the present invention uses rotating three-dimensional magnetic field microfluidics technology for nucleic acid analysis, which can realize rapid and efficient mixing of large-volume samples and magnetic beads, so that low-concentration samples can be enriched by increasing the injection volume, and higher detection efficiency can be obtained. sensitivity;

2. The present invention integrates a one-dimensional mobile magnetic field control module and a flat reaction cell, and only needs one-dimensional reciprocating motion to realize efficient mixing and reaction of magnetic beads and reagents. The chip structure and control method are relatively simple, and are suitable for multi-step magnetic beads The operation process is simple, and it is easy to integrate other functional units to build an automated, portable, and low-cost nucleic acid micro-analysis system.

Description of drawings

Figure 1 is a schematic diagram of the microfluidic chip system structure for full nucleic acid analysis;

Fig. 2 is a schematic structural diagram of a microfluidic chip;

Fig. 3 is a schematic diagram of the mixing principle of magnetic beads in a rotating three-dimensional magnetic field;

Figure 4 is a schematic structural diagram of a three-channel/four-fluid pool microfluidic chip for full nucleic acid analysis;

Figure 5 is a schematic diagram of the structure of a two-channel/seven-fluid pool microfluidic chip for full nucleic acid analysis;

Figure 6 The effect of three-dimensional mixing time of nucleic acid extraction on the _CT value of the real-time fluorescent PCR amplification curve.

In the figure: 1, rotating three-dimensional magnetic field control module, 11, upper circular plate, 12, upper circular plate magnet, 13, lower circular plate, 14, lower circular plate magnet, 15, driving motor, 16, rotating shaft, 17, rotating shaft base ;2. One-dimensional mobile magnetic field control module, 21. Rectangular permanent magnet fixed component, 22. Microfluidic chip mobile control component; 3. Temperature control module; 4. Signal detection module; 5. Microfluidic chip, 51. Three-dimensional Mixing tank, 52, reaction tank, 53, liquid feeding hole, 54, isolation tank.

Detailed ways

The following non-limiting examples can enable those skilled in the art to understand the present invention more fully, but do not limit the present invention in any way. In the schematic diagram of the microfluidic chip and device of the present invention, the structural dimensions are not marked, which mainly reflects the structural structure of the microfluidic chip and device. During actual production and use, the structural ratio and size can be adjusted as needed.

Example 1

The present invention will be further described below in conjunction with the accompanying drawings. Fig. 1 is a structural block diagram of the nucleic acid full-analysis microfluidic chip system of the present invention, Fig. 2 is a structural diagram of the microfluidic chip, and Fig. 3 is a schematic diagram of the principle of rotating three-dimensional magnetic field magnetic beads mixing; control chip 5 and a portable device, wherein the portable device is composed of a rotating three-dimensional magnetic field control module 1, a one-dimensional moving magnetic field control module 2, a temperature control module 3 and a signal detection module 4.

Wherein, the rotating three-dimensional magnetic field control module 1 of the present invention is composed of an upper circular plate 11 , an upper circular plate magnet 12 , a lower circular plate 13 , a lower circular plate magnet 14 , a driving motor 15 , a rotating shaft 16 and a rotating shaft base 17 . Specifically, the upper circular plate 11 and the lower circular plate 13 are arranged parallel to each other horizontally, and the upper circular plate 11 and the lower circular plate 13 are provided with a rotating shaft 16, and the rotating shaft 16 is arranged at a position deviated from the center of the circle and vertically Through the two circular plates; the top of the rotating shaft 16 is provided with a driving motor 15 , and the bottom of the rotating shaft 16 is provided with a rotating shaft base 17 , and the upper circular plate 11 and the lower circular plate 13 are driven by the driving motor 15 through the rotating shaft 16 . The diameters of the upper circular plate 11 and the lower circular plate 13 used in this embodiment are both 45 mm, and the thickness is 3 mm. The eccentricity of the rotating shaft 16 is 7.5 mm; 6 upper disc magnets, 12, the lower disc 13 edge is provided with 6 lower disc magnets 14 with the same shape and evenly arranged, the long sides of the upper disc magnet 12 and the lower disc magnet 14 Perpendicular to the rotating shaft 6, the upper disc magnet 12 and the lower disc magnet 14 are 8.5 mm long, 1.5 mm wide and 5 mm high, and the upper disc magnet 12 and the lower disc magnet 14 are NdFeB permanent magnets, And the upper disc magnet 12 and the lower disc magnet 14 are staggered vertically. The microfluidic chip 5 used in the embodiment of the present invention has a three-dimensional mixing pool 51 located at one end of the microfluidic chip 5, which is a closed structure with unlimited shape and length, and a depth > 1 mm. In the magnetic field region between the circular plates 13.

The one-dimensional mobile magnetic field control module 2 includes a microfluidic chip 5, a rectangular permanent magnet fixing component 21 and a microfluidic chip mobile control component 22, and one end of the microfluidic chip 5 is provided with a three-dimensional mixing pool 51, so The three-dimensional mixing pool 51 is located on the lower circular plate magnet 14 of the lower circular plate 13, the other end of the microfluidic chip 5 is provided with a microfluidic chip mobile control assembly 22, and the microfluidic chip mobile control assembly 22 is provided. The lower end of the microfluidic chip 5 at one end of 22 is provided with a rectangular permanent magnet fixing component 21 adjacent to the side of the microfluidic chip mobile control component 22, and the other side of the rectangular permanent magnet fixing component 21 is provided with a temperature control module 3, The temperature control module 3 is located at the lower end of the microfluidic chip 5, and is close to the bottom plate of the microfluidic chip. The upper end of the microfluidic chip 5 is provided with a signal detection module 4, and the signal detection module 4 is located above the temperature control module 3. .

As shown in Figure 2, there are at least two channels on the microfluidic chip 5, the channels are arranged in parallel, and each channel is provided with a three-dimensional mixing pool 51 and at least two reaction pools 52 in sequence, An isolation pool 54 is provided between the three-dimensional mixing pool 51 and the reaction pool 52, and an isolation pool 54 is provided between the adjacent reaction pools 52. The three-dimensional mixing pool 51, the reaction pool 52 and the isolation pool 54 are all A liquid filling hole 53 is provided, one side of the passage is a three-dimensional mixing pool 51 , and the other side of the passage is an isolation pool 54 . The depth of the three-dimensional mixing tank 51 in this embodiment is 2 mm, the reaction tank 52 is flat, and the depth is 0.4 mm, and the volume ratio of the three-dimensional mixing tank 51 and the reaction tank 52 is 10:1.

The temperature control area of the temperature control module 3 covers the desired position of the reaction pool, and the temperature control module 3 includes a temperature control element and a control circuit.

The detection area of the signal detection module 4 covers the required position of the reaction pool, and the signal detection module 4 includes a signal acquisition unit and a data processing unit.

The principle of mixing magnetic beads in a rotating three-dimensional magnetic field is described in detail below in conjunction with the schematic diagram in Figure 3:

Place the three-dimensional mixing pool 51 containing nucleic acid samples and magnetic beads in the magnetic field area between the upper circular plate 11 and the lower circular plate 13 of the rotating three-dimensional magnetic field control module 1 of the present invention, after starting the drive motor 15, the upper and lower The lower circular plate starts to rotate around the rotating shaft 16 at the same time; when the circular plate is rotated, the magnets embedded in the circular plate generate a periodic and gradual three-dimensional magnetic field between the two layers of circular plates, thereby controlling the three-dimensional mixing pool 51 located in this area. The magnetic beads do three-dimensional S-shaped movement, thereby realizing full contact, efficient mixing and rapid reaction of magnetic beads and nucleic acid samples, and significantly improving the binding rate of magnetic beads and nucleic acid molecules. After 1/2 cycle, the magnetic beads move from one end of the three-dimensional mixing pool 51 to the other end in an S-shaped space (see FIG. 3, a), and its periodic motion track is shown in FIG. 3(b).

Example 2

The microfluidic chip described in this example is schematically shown in Figure 4. It is a three-channel/four-fluid pool microfluidic chip, which contains three nucleic acid analysis channels, and each channel contains four reaction pools, each of which is a three-dimensional mixing pool. pool, two cleaning pools and one elution/amplification pool, and the isolation pool between the reaction pools; among them, the depth of the three-dimensional mixing pool is 2mm, the width is 4.5mm, and the depth of the cleaning pool and the elution/amplification pool is 0.4mm , with a width of 2.25mm.

As can be seen from Figure 4, the method for performing full nucleic acid analysis using the chip and device described in the embodiment of the present invention includes the following steps:

1) Microfluidic chip preparation: Add general-purpose magnetic silica beads and their buffers (such as 6M guanidine hydrochloride, 10mM Tris-1mM EDTA buffer, pH 6.1) and nucleic acid samples to the three-dimensional mixing pool, and add washing buffer to the washing pool solution (such as 75-80% ethanol), add nucleic acid amplification/detection reagents (such as real-time fluorescent PCR reaction solution except the template) to the elution/amplification pool, add mineral oil as the spacer medium to the spacer pool, and then seal each The filling hole of the liquid pool;

2) Nucleic acid extraction based on a microfluidic chip: place the three-dimensional mixing pool (51) completely in the rotating three-dimensional magnetic field control area, start the drive motor 15, and drive the magnetic beads in the three-dimensional mixing pool to perform a three-dimensional movement, so that the magnetic beads and the sample are fully mixed. After contacting and mixing, the nucleic acid molecules in the solution are efficiently adsorbed to the surface of the magnetic beads; then, the driving motor 15 is turned off, and the microfluidic chip movement control component 22 is activated, so that the magnetic beads leave the three-dimensional mixing pool and enter the first cleaning pool through the isolation pool and perform one-dimensional reciprocating motion to fully clean the impurities adsorbed on the surface of the magnetic beads; then control the microfluidic chip movement control component 22 to make the magnetic beads enter the second cleaning pool and perform one-dimensional reciprocating motion to further clean the magnetic beads Impurities adsorbed on the bead surface.

3) Nucleic acid amplification and detection based on the microfluidic chip: control the mobile control component 22 of the microfluidic chip, so that the magnetic beads enter the elution/amplification pool and perform one-dimensional reciprocating motion, and the nucleic acid molecules adsorbed on the surface of the magnetic beads Fully eluted into the amplification reaction solution; finally, move the elution/amplification pool to the temperature control module 3 area, start the temperature control module 3 and the signal detection module 4, start the nucleic acid amplification and detection process, and complete the automatic nucleic acid full-blown Analysis process.

Example 3

The microfluidic chip described in this example is schematically shown in Figure 5. It is a two-channel/seven-fluid pool microfluidic chip, which contains two nucleic acid analysis channels, and each channel contains seven reaction pools, each of which is a three-dimensional mixing pool. pool, two cleaning pools, one amplification pool, the other two cleaning pools and one detection pool, and an isolation pool between the reaction pools; among them, the depth of the cleaning pool, amplification pool and detection pool is less than 0.5mm, and the width is smaller than that of the three-dimensional mixing pool. 1/2 of the pool width.

As can be seen from Figure 5, the method for performing full nucleic acid analysis using the chip and device described in the embodiment of the present invention includes the following steps:

1) Microfluidic chip preparation: add DNA capture probe modified magnetic beads and its buffer and nucleic acid samples into the three-dimensional mixing pool, add washing buffer (such as 75-80% ethanol) into the cleaning pool, and add Add nucleic acid amplification reagents (such as PCR reaction solution except the template), pre-fix the DNA detection probe microarray in the detection pool, add spacer medium in the isolation pool, and then seal the liquid filling holes of each liquid pool;

2) Nucleic acid purification based on the microfluidic chip: fix the microfluidic chip (5) on the bracket and adjust it so that the three-dimensional mixing pool (51) is completely located in the rotating three-dimensional magnetic field control area, start the driving motor 15, and drive the three-dimensional mixing pool The three-dimensional movement of the magnetic beads realizes that the magnetic beads and the sample are fully contacted and mixed, and the nucleic acid molecules containing specific sequences in the solution are efficiently combined with the DNA probes on the surface of the magnetic beads; then, the drive motor 15 is turned off, and the movement control of the microfluidic chip is started. Component 22, making the magnetic beads leave the three-dimensional mixing pool, pass through the isolation pool and enter the first cleaning pool and perform one-dimensional reciprocating motion to fully clean the impurities on the surface of the magnetic beads and non-specifically bound nucleic acid molecules; then control the microfluidic chip The control assembly 22 is moved to make the magnetic beads enter the second cleaning pool and perform one-dimensional reciprocating motion, so as to further clean the impurities and non-specifically bound nucleic acid molecules on the surface of the magnetic beads.

3) Nucleic acid amplification based on the microfluidic chip: control the mobile control component 22 of the microfluidic chip, make the magnetic beads enter the amplification pool and move the amplification pool to the temperature control module 3 area, start the temperature control module 3 to start the nucleic acid Amplification process.

4) Nucleic acid multi-indicator detection based on microfluidic chip DNA microarray: After the amplification is completed, control the microfluidic chip mobile control component 22 to efficiently combine the magnetic beads with the reaction products in the amplification pool, and then make the magnetic beads sequentially Enter the following two cleaning pools to fully clean the residual impurities and non-specifically bound nucleic acid molecules on the surface of the magnetic beads; finally, make the magnetic beads enter the detection pool and perform one-dimensional reciprocating motion, and start the temperature control module 3 at the same time to start the nucleic acid hybridization reaction Process: After the hybridization reaction is completed, the signal detection module 4 is started to obtain the nucleic acid multi-indicator detection results and complete the automated nucleic acid full analysis process.

Example 4

The microfluidic chip described in this example is schematically shown in Figure 4. A commercialized magnetic bead method bacterial genomic DNA extraction kit (AU2001, Beijing Biotech Biotechnology Co., Ltd.) was used, and the chip described in the embodiment of the present invention and The device investigates the effect of three-dimensional mixing time on the nucleic acid extraction effect, including the following steps:

1) Nucleic acid sample preparation: centrifuge 1.5mL Vibrio harveylius solution at 12000r/min for 3min, remove the supernatant; add 1mL of TE buffer and mix well, boil the resuspended bacteria solution at 100°C for 5-10min, Then centrifuge at 12000r/min for 3min, take the supernatant, store at -20°C or use immediately;

2) Microfluidic chip preparation: Add magnetic bead binding solution CB, mixed magnetic bead solution and lysed bacteria solution to the three-dimensional mixing pool, add rinsing solution WB to the cleaning pool, and add elution solution to the elution/amplification pool. Buffer TE, add mineral oil to the spacer;

3) Microfluidic chip-based nucleic acid extraction: fix the microfluidic chip 5 on the bracket and adjust the three-dimensional mixing pool 51 to be completely located in the rotating three-dimensional magnetic field control area, then set the experimental parameters, start the portable device, and start chip nucleic acid extraction experiment. In order to investigate the effect of three-dimensional mixing conditions on the binding efficiency of magnetic beads and DNA molecules, different three-dimensional mixing times were set (0, 1, 5, and 9 min, respectively).

4) Detection of chip nucleic acid extraction results: After the chip nucleic acid extraction process is completed, draw all the solutions from the liquid filling hole of the elution pool into the centrifuge tube, and perform real-time fluorescent PCR detection _. The influence of the value is shown in Figure 6, which shows that the DNA sample extracted by the chip can be directly used for PCR amplification, and within 0-9min, the longer the three-dimensional mixing time, the higher the extracted DNA concentration, which proves that the three-dimensional mixing of magnetic beads and samples has a positive effect. It helps to improve the binding efficiency of DNA molecules and magnetic beads in the sample.

Claims (9)

1. a kind of nucleic acid complete analysis micro-fluidic chip system, it is characterised in that: including rotated three dimensional magnetic field control module (1), one Tie up shifting magnetic field control module (2), temperature control modules (3), signal detection module (4) and micro-fluidic chip (5), in which:

The rotated three dimensional magnetic field control module (1) include upper plectane (11), upper plectane magnet (12), lower plectane (13), under Plectane magnet (14), driving motor (15), shaft (16) and shaft base (17)；The upper plectane (11) and lower plectane (13) are Corresponding horizontal parallel setting, the upper plectane (11) and lower plectane (13) are equipped with shaft (16), and the shaft (16) is set to Deviate the position in the center of circle and extends vertically through in upper plectane (11) and lower plectane (13)；Shaft (16) top is equipped with driving motor (15), shaft (16) bottom end is equipped with shaft base (17), and the upper plectane (11) and lower plectane (13) pass through shaft (16) It is driven by driving motor (15)；It is identical, uniform that the upper plectane (11) with the edge of lower plectane (13) is successively respectively equipped with shape The multiple upper plectane magnets (12) and lower plectane magnet (14) of arrangement, the upper plectane magnet (12) and lower plectane magnet (14) Long side is vertical with shaft (16), and the upper plectane magnet (12) and lower plectane magnet (14) are mutually handed on upright position It is wrong；

The one-dimensional movement magnetic field control module (2) include micro-fluidic chip (5), rectangle permanent magnet fixation kit (21) and Micro-fluidic chip is mobile control assembly (22)；One end of the micro-fluidic chip (5) is equipped with three-dimensional hybrid pond (51), the three-dimensional Mixing pit (51) is in the field region between upper plectane (11) and lower plectane (13), the micro-fluidic chip (5) it is another The side at end is equipped with micro-fluidic chip movement control assembly (22), and the lower end of the micro-fluidic chip (5) is close in micro-fluidic core Mobile control assembly (22) side of piece is equipped with rectangle permanent magnet fixation kit (21), the rectangle permanent magnet fixation kit (21) the other side is equipped with temperature control modules (3), and the temperature control modules (3) are close to micro-fluidic chip (5) lower end, described Micro-fluidic chip (5) upper end is equipped with signal detection module (4), and the signal detection module (4) is located on temperature control modules (3) Side.

2. nucleic acid complete analysis micro-fluidic chip system according to claim 1, it is characterised in that: the micro-fluidic chip (5) at least two channels on, the channel are parallel arrangement, are successively arranged a three-dimensional hybrid pond on each channel (51) and at least two reaction tanks (52) isolation tank (54), are equipped between the three-dimensional hybrid pond (51) and reaction tank (52), institute It states and is equipped with isolation tank (54) between adjacent reaction tank (52), the three-dimensional hybrid pond (51), reaction tank (52) and isolation tank (54) it is equipped on liquid filling hole (53), the side endpoint in the channel is three-dimensional hybrid pond (51), another side in the channel Point is isolation tank (54).

3. nucleic acid complete analysis micro-fluidic chip system according to claim 1, it is characterised in that: the three-dimensional hybrid pond It (51) is enclosed construction, depth > 1mm, the reaction tank (52) is platypelloid type, depth < 0.5mm, the three-dimensional hybrid pond (51) with the volume ratio > 4:1 of reaction tank (52).

4. nucleic acid complete analysis micro-fluidic chip system according to claim 1, it is characterised in that: the temperature control modules (3) temperature control region overlay reaction tank (52), the temperature control modules (3) include temperature control component and control circuit.

5. nucleic acid complete analysis micro-fluidic chip system according to claim 1, it is characterised in that: the signal detection module (4) detection zone covers reaction tank (52), and the signal detection module (4) includes signal acquisition unit and data processing list Member.

6. the application method of nucleic acid complete analysis micro-fluidic chip system according to claim 1, it is characterised in that: including following Step:

1. the modified magnetic bead in surface and nucleic acid samples to be detected are added in three-dimensional hybrid pond (51), it is added in reaction tank (52) corresponding Reagent, blank medium is added in isolation tank (54), the liquid filling hole of each liquid pool is then closed using sealed membrane；

2. three-dimensional hybrid pond (51) are fully seated at rotated three dimensional magnetic field control area, start rotated three dimensional magnetic field control module (1), it is rotated by plectane (11) in drive shaft (16) drive and lower plectane (13), come into full contact with magnetic bead and nucleic acid samples, Mixing；

3. step 2. after the completion of, close rotated three dimensional magnetic field control module (1), the mobile control assembly of starting micro-fluidic chip (22), magnetic bead is made to leave three-dimensional hybrid pond and enter in next reaction tank by isolation tank；

4. step 3. after the completion of, control micro-fluidic chip is mobile control assembly (22), carries out magnetic bead in reaction tank one-dimensional past Multiple movement is realized magnetic bead and being sufficiently mixed of reagent in reaction tank, is reacted；

5. step 4. after the completion of, control micro-fluidic chip is mobile control assembly (22), and magnetic bead is made to sequentially enter subsequent reactions pond, suitable It is secondary to complete subsequent mixing, reaction process, and reacted and detected in last reaction tank, to realize nucleic acid complete analysis stream Journey.

7. application method according to claim 6, it is characterised in that: the step 1. in the modified magnetic bead in surface include logical With property magnetism silica bead, specificity DNA probing needle modified magnetic bead, immunomagnetic beads.

8. application method according to claim 6, it is characterised in that: the step 1. in the reagent that is added into reaction tank Including cleaning buffer solution, elution buffer, nucleic acid amplification agents, detection reagent.

9. application method according to claim 6, it is characterised in that: the step 1. in spacer medium be and water not phase Another flow media held, another flow media incompatible with water includes air, mineral oil.