CN113150979A - Portable nucleic acid detector based on micropore array chip - Google Patents
Portable nucleic acid detector based on micropore array chip Download PDFInfo
- Publication number
- CN113150979A CN113150979A CN202011642787.3A CN202011642787A CN113150979A CN 113150979 A CN113150979 A CN 113150979A CN 202011642787 A CN202011642787 A CN 202011642787A CN 113150979 A CN113150979 A CN 113150979A
- Authority
- CN
- China
- Prior art keywords
- nucleic acid
- array chip
- detection
- portable
- acid detector
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
Landscapes
- Chemical & Material Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Health & Medical Sciences (AREA)
- Biophysics (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Immunology (AREA)
- Microbiology (AREA)
- Molecular Biology (AREA)
- Analytical Chemistry (AREA)
- Physics & Mathematics (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
一种基于微孔阵列芯片的便携式核酸检测仪，包括：核酸提取模块、核酸扩增模块和检测模块；其中，所述核酸扩增模块包括微孔阵列芯片放置槽和温控组件，所述微孔阵列芯片放置槽用于放置微孔阵列芯片，所述温控组件位于所述微孔阵列芯片放置槽的下方。本发明的便携式核酸检测仪可以同时对多样本多指标的样本进行同时核酸提取处理及扩增检测，大大缩短了检测时间，可实现现场快速检测。
A portable nucleic acid detector based on a microporous array chip, comprising: a nucleic acid extraction module, a nucleic acid amplification module and a detection module; wherein the nucleic acid amplification module comprises a microporous array chip placement groove and a temperature control assembly, the microporous array chip The hole array chip placement groove is used for placing the micropore array chip, and the temperature control component is located below the micropore array chip placement groove. The portable nucleic acid detector of the present invention can simultaneously perform nucleic acid extraction processing and amplification detection on samples with multiple samples and multiple indicators, greatly shortens the detection time, and can realize rapid detection on site.
Description
Technical Field
The invention relates to a high-throughput rapid portable nucleic acid detector, which integrates the functions of automatic nucleic acid extraction, isothermal amplification detection and quantitative reading, can be used for field detection of various pathogens including new coronavirus, malaria and the like, and can really realize sample in-and-out. Based on the self-developed high-flux micropore array chip, the portable detector can realize high-flux synchronous detection, and the detection result can be quantitatively displayed on a liquid crystal display screen of the portable detector.
Background
A new type of coronavirus pneumonia (coronaviral disease 2019, COVID-19) caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) rapidly breaks out in the world. Great efforts have been made in the field of public health, and technological progress has been made, but the work of prevention and control of new coronary pneumonia epidemic is still a major challenge facing human beings from the global view. The rapid and accurate diagnosis of COVID-19 is of great significance to control virus infection and carry out early treatment.
According to different detection principles, the detection methods of the new coronavirus which are commercially used at present are mainly divided into two types: an immunoassay-based serum antibody indirect detection method and a method for directly detecting the new coronavirus nucleic acid based on a throat swab sample. Immunodetection techniques such as enzyme-linked immunosorbent assay (ELISA), immunofluorescence, and lateral flow immunoassay test strips (LFI) are directed against novel coronavirus antibodies produced in blood, including immunoglobulin M (IgM) and immunoglobulin G (IgG). The method has low detection cost and low requirements on detection environment and detection equipment, and can realize rapid qualitative detection on site. However, since the generation of virus antibodies usually takes 3-7 days, the method is more suitable for the prognosis detection of new coronary pneumonia. The nucleic acid detection technology based on molecular diagnosis comprises the amplification technology of the classical common Polymerase Chain Reaction (PCR) and the emerging constant temperature amplification technology, is a gold standard for detecting the new coronavirus due to the characteristics of good specificity, high accuracy, strong sensitivity and the like, and plays an important role in early diagnosis of the new coronavirus and investigation of suspected patients.
However, in the detection of various infectious pathogens including the novel coronavirus, the conventional nucleic acid detection technique needs to be performed by a special operator in a laboratory with a biosafety level of two or more. The main detection process comprises the steps of inactivation of a throat swab sample, enrichment and extraction of a nucleic acid sample based on a magnetic bead method and nucleic acid amplification detection, has high requirements on environment, complex operation and expensive equipment, limits the application of the detection in large-scale screening and on-site rapid detection, and greatly promotes the development of molecular diagnosis instant detection equipment.
At present, there are reported developed or marketed molecular diagnosis instant detection devices, such as a nucleic acid detector (CN 107043697) integrating a microfluidic chip and a constant temperature amplification detection technology, which uses the microfluidic chip as a detection consumable, has high cost, and can only perform single-flux detection; or a portable nucleic acid detection kit (CN 207276621) of a mobile PCR detection laboratory, has high instrument cost and is not suitable for field rapid detection in remote areas. Therefore, the low-cost portable nucleic acid detector capable of simultaneously integrating automatic nucleic acid extraction, high-throughput detection and quantitative reading is few in domestic markets at present, and has important significance for on-site rapid detection and large-scale screening of various pathogens including new coronavirus.
Disclosure of Invention
In order to solve the problems in the prior art, the invention provides a portable nucleic acid detector based on a micropore array chip, which can realize high-throughput detection and reduce the cost.
In order to achieve the purpose, the invention adopts the following technical scheme:
a portable nucleic acid detector based on a micro-well array chip, comprising: a nucleic acid extraction module, a nucleic acid amplification module and a detection module; the nucleic acid amplification module comprises a micropore array chip placing groove and a temperature control component, wherein the micropore array chip placing groove is used for placing a micropore array chip, and the temperature control component is positioned below the micropore array chip placing groove.
In some embodiments, the microwell array chip has a microwell upper diameter that is smaller than a microwell lower diameter.
In some embodiments, the microwell array chip has a thickness of 2cm to 5cm (e.g., 2.5mm, 3.0mm, 3.5mm, 4.0mm, or 4.5 mm).
In some embodiments, the upper diameter of the microwells is between 500 μm and 2mm (e.g., 800 μm, 1.0mm, 1.2mm, 1.5mm, or 1.8mm) and the lower diameter of the microwells is between 1mm and 5mm (e.g., 1.5mm, 2.0mm, 2.5mm, 3.0mm, 3.5mm, 4.0mm, or 4.5 mm).
In some embodiments, after sample application, the microwell array chip is closed with a sealing membrane.
In some embodiments, the detection module comprises an image capture unit positioned over the microwell array chip.
In some embodiments, the portable nucleic acid testing instrument includes an upper cover, and the image capturing unit is disposed inside the upper cover.
In some embodiments, the detection module further comprises a display screen for detecting the nucleic acid detection result.
In some embodiments, the nucleic acid amplification module performs nucleic acid amplification using a loop-mediated isothermal amplification technique.
In some embodiments, the portable nucleic acid testing apparatus further comprises a sample storage module for storing a sample to be tested.
Compared with the prior art, the portable nucleic acid detector based on the micropore array chip has the beneficial effects that:
(1) the portable nucleic acid detector and the matched high-flux micropore array chip can simultaneously carry out simultaneous nucleic acid extraction treatment and constant-temperature amplification detection on a multi-sample and multi-index sample, and can complete sample RNA extraction, magnetic bead enrichment, isothermal detection and data analysis within 35 minutes, thereby greatly shortening the detection time;
(2) the portable nucleic acid detector integrates the full-automatic nucleic acid extraction function and the detection function, can realize on-site rapid detection without other auxiliary equipment, automatically and quantitatively analyzes the detection result, is readable by naked eyes, and has no special professional requirements on operators.
(3) The portable nucleic acid detector has the advantages of simple structure, integration of the sample pretreatment system, the micropore array chip and the constant-temperature amplification nucleic acid detection equipment, reduction of operation steps, realization of integration of nucleic acid extraction, amplification and detection, small volume, light weight and convenience in carrying, and is high-efficiency high-throughput chip type constant-temperature amplification nucleic acid detection equipment.
(4) The portable nucleic acid detector and the high-flux micropore array chip have low preparation cost, wherein the processing cost of the portable nucleic acid detector is 1 ten thousand yuan per platform, and the cost of the micropore array chip is 10 yuan per chip, thereby greatly reducing the preparation and detection cost.
Drawings
The drawings are only for purposes of illustrating and explaining the present invention and are not to be construed as limiting the scope of the present invention. Wherein:
FIG. 1 is a schematic view showing the structure of a portable nucleic acid detecting apparatus according to an embodiment of the present invention;
FIG. 2 is a side view of a portable nucleic acid detecting apparatus according to an embodiment of the present invention;
FIG. 3 is an external view of a portable nucleic acid detecting apparatus according to an embodiment of the present invention;
FIG. 4 is a schematic diagram of a circuit structure of the portable detector according to an embodiment of the present invention;
FIG. 5 illustrates a minimum detection limit verification for a detection system in an embodiment of the present invention;
FIG. 6 is a comparison of detection times of a microwell array chip and a commercial octal tube in an example of the present invention.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is further described in detail below with reference to the accompanying drawings in combination with specific embodiments.
In the description of the present invention, reference to "one embodiment" means that a particular feature, structure, or parameter, step, or the like described in the embodiment is included in at least one embodiment according to the present invention. Thus, appearances of the phrases such as "in one embodiment," "in one embodiment," and the like in this specification are not necessarily all referring to the same embodiment, nor are other phrases such as "in another embodiment," "in a different embodiment," and "in a different embodiment" unless expressly stated otherwise. Those of skill in the art will understand that the particular features, structures or parameters, steps, etc., disclosed in one or more embodiments of the present description may be combined in any suitable manner.
In the description of the present invention, it is to be noted that the terms "connected" and "connected" are to be interpreted broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected, unless explicitly stated or limited otherwise; they may be directly connected or indirectly connected through an intermediate. The specific meaning of the above terms in the present invention can be understood as appropriate by those of ordinary skill in the art.
The portable nucleic acid detector of the invention takes a loop-mediated isothermal amplification technology as a detection technology, combines a high-flux micropore array chip, and designs a throat swab sample storage module, a nucleic acid extraction module, a nucleic acid amplification module with small temperature control area and low energy consumption and a detection module. The portable nucleic acid detector disclosed by the invention does not need the assistance of other laboratory equipment, is simple to operate by a user, realizes the detection of multiple indexes and multiple samples on the basis of portability, can directly display the detection result on a liquid crystal display screen, and is very suitable for the fields of epidemic detection inspection, field quick detection and the like.
As shown in FIGS. 1 and 2, the portable nucleic acid detecting instrument of the present invention comprises an upper cover 1 and a main body 2, and the upper cover 1 is provided with a detecting module including a display 3 (e.g., a liquid crystal display) at the top of the outer side of the upper cover and an image pickup unit, e.g., a camera module 4, at the inner side. Be provided with pharynx swab sample storage module 5, nucleic acid extraction module 6 and nucleic acid amplification module on the main part 2, pharynx swab sample storage module 5 and nucleic acid extraction module 6 are located around the nucleic acid amplification module, and pharynx swab sample storage module 5 includes a plurality of slots, can place containers such as centrifuging tube that are used for preserving the sample, based on nucleic acid preservation solution, can preserve 3 ~ 16 different pharynx swab samples simultaneously.
According to an embodiment of the present invention, the nucleic acid extraction module 6 includes a plurality of slots for accommodating containers such as centrifuge tubes for containing nucleic acid lysates. The nucleic acid extraction module 6 may also include a multichannel pipettor 7. The nucleic acid amplification module comprises a micropore array chip placing groove 8 and a temperature control component 9, wherein the micropore array chip placing groove 8 is used for placing a micropore array chip for nucleic acid amplification, and the temperature control component 9 is positioned below the micropore array chip placing groove 8 and used for heating the micropore array chip.
In the embodiment of the invention, the extraction of nucleic acid can be realized by combining a magnetic bead method and a numerical control manipulator. And controlling the numerical control mechanical arm, moving the numerical control mechanical arm to the sample storage module 5, sucking a certain amount of samples, and automatically transferring the samples to a nucleic acid lysate for cracking, wherein the amount of the sucked samples can be 200-1000 mu L. The nucleic acid extraction module can simultaneously process 3-16 samples. And (3) enriching the nucleic acid sample after the cracking by adopting magnetic beads. And finally, automatically sucking a certain amount of nucleic acid lysate, and transferring the nucleic acid lysate to the micro-pore array chip, wherein the amount of the sucked sample can be 5-20 mu L.
The working principle of the portable nucleic acid detector in the embodiment of the invention is a loop-mediated isothermal amplification technology, and the high-throughput micropore array chip based on the nucleic acid amplification module realizes the simultaneous isothermal amplification detection of 3-16 samples. In the detection process, a color developing agent is added, wherein the color developing agent comprises but is not limited to color indication and fluorescence reaction, a color image of a detection area can be obtained by using the camera assembly 4, the color or fluorescence intensity is automatically extracted and distinguished by calibrating a comparison area, an interpretation result is given after automatic analysis, and finally the interpretation result is displayed on the display screen 3.
Compared with the common PCR technology which requires variable temperature for amplification, the loop-mediated isothermal amplification can realize 10 times of specific nucleic acid sequence by using strand displacement DNA polymerase within 15-60 minutes under the condition of isothermal temperature (60-65℃)9～1010Amplification of the fold is rapid. Based on the technology, specific primers are designed aiming at specific gene regions of the new coronavirus, and plasmids of related genes and clinical practical samples are obtainedTests were performed to verify the specificity and sensitivity of the primers.
In order to carry out low-cost and high-flux detection on samples such as novel coronavirus and the like, the invention designs a high-flux micropore array chip suitable for the portable nucleic acid detector.
In some embodiments, the microwells of the microwell array chip are fabricated by etching a substrate made of polymethyl methacrylate or the like using a laser etching technique. The microporous structure can adopt a conical structure, namely the upper diameter of the micropores is smaller than the lower diameter of the micropores, so that the evaporation of a solution during isothermal amplification reaction can be effectively reduced, the possibility of nucleic acid aerogel production is reduced, and false positive results caused by cross contamination are avoided. After the sample adding is finished, the upper surface of the chip is sealed by using the single-sided sealing film, so that the mutual crosstalk among different detection sites is avoided, and the detection accuracy is effectively improved.
The detection flux of the micropore array chip can be adjusted according to actual requirements, and 3-16 samples can be detected simultaneously. A positive control (P) and a negative control (N) can be designed in the micropore array chip, and the positive control avoids the detection result of false negative; the negative control avoids 'false positive' results caused by environmental pollution.
The size of the volume of the microwell is designed and adjusted according to the amount of sample to be detected and the volume of reagents required for amplification, and in some embodiments, the amount of sample carrying a nucleic acid detection reagent is 20 to 50. mu.L, the thickness of the chip may be 2 to 5cm (e.g., 2.5mm, 3.0mm, 3.5mm, 4.0mm, or 4.5mm), the upper diameter of the microwell may be 500 to 2mm (e.g., 800. mu.m, 1.0mm, 1.2mm, 1.5mm, or 1.8mm), and the lower diameter of the microwell may be 1 to 5mm (e.g., 1.5mm, 2.0mm, 2.5mm, 3.0mm, 3.51nm, 4.0mm, or 4.5 mm).
As shown in FIG. 3, the micro well array chip can be placed in a chip placement slot of a portable nucleic acid detector, the temperature control assembly 9 heats the micro well array chip and obtains an image of an amplification detection area, and software on the main control board automatically analyzes the image and judges a detection result to be displayed on the display screen 3. At present, 130 clinical sample tests are carried out in western China hospital, the detection sensitivity is 95.4%, and the detection specificity is 95.35%.
As shown in FIG. 4, the circuit structure of the portable nucleic acid detecting apparatus of the present invention includes a main control board, a liquid crystal display, a main control circuit, a camera module, a temperature control device, a control circuit board, a power supply, a motor and driver module, a fan, a power interface, and a power switch. The main control circuit is respectively connected with the temperature control element, the camera shooting assembly, the motor and driver assembly and the main control computer and is controlled by the control circuit board. The main control computer is connected with the power supply and is connected with the camera shooting assembly.
As shown in FIG. 3, in some embodiments, the portable nucleic acid detecting instrument of the present invention employs a flip design, wherein the flip not only serves to seal the instrument and shield external dust and foreign matter, but also has a main control computer mounted therein, and the main control computer is connected to the power supply and the camera module, and is sealed by four screws. The camera shooting assembly is positioned above the micropore array chip, can acquire a color image of the amplification reaction area and feed the image back to the main control board, and software on the main control board automatically analyzes the image and judges a detection result to be displayed on a display screen; the display screen may be a touch screen, and is configured to respond to a touch operation of an operator and output a signal.
The inside a plurality of baffles that are provided with of main part for put the constant head tank of consumptive material, including the centrifuging tube slot of sample storage module and nucleic acid extraction module, and the micro-control array chip of nucleic acid amplification module place the groove. The bottom of the box body is provided with a control circuit board which is responsible for the operation of power management, master control board communication, temperature control components, a heating control circuit board, a camera component and a nucleic acid extraction module of the equipment. The temperature control component is used for heating and controlling the constant temperature of the micropore array chip, and the temperature can be 30-100 ℃. Other accessories such as fans are used to dissipate heat from the devices inside the apparatus; the power interface is used for supplying power to the equipment through the external power adapter; the power switch is used for controlling the whole equipment to be switched on and off.
The fixed platform, the support, the camera shooting component fixing frame, the heat conducting plate, the motor frame and the heating pipe are made of aluminum alloy materials. The surface treatment of the heat conducting plate adopts natural color oxidation, the surface has a frosted effect, and the quality of a detected image can be improved by the clean and uniform fine frosted effect. The chip placing groove, the heat conducting plate fixing frame, the heating film cover plate and the battery pressing plate are made of Polyformaldehyde (POM) materials, wherein the chip placing groove and the heat conducting plate fixing frame need to be made of black POM materials, and the surface of the chip placing groove needs to be flat and smooth due to the fact that the appearance of the chip placing groove is visible. In one embodiment, the portable detector has dimensions of 25 × 15 × 15 cm.
In the detection process, the collected throat swab sample is stored in a sample storage module, and the sample is inactivated at 56 ℃ for 5 min.
The RNA in a detection sample is extracted based on an amplification and detection module in a portable nucleic acid detector, and the specific extraction steps are as follows: using a numerical control robot arm, 800ul of the sample solution was automatically transferred and added to 20ul of the extracting solution containing the nucleolytic enzyme (magnetic bead method), and the nucleic acid was extracted. Setting equipment operation parameters: mixing the magnetic beads for 1 min; cracking for 5 min; washing I: 1 min; washing II: 1 min; nucleic acid elution: 3 min; and (4) magnetic bead suction and abandoning: for 1 min. After the automatic extraction is finished, RNA is collected for subsequent experiments.
Then the extracted RNA is used as a template to be added into a reaction system of the loop-mediated isothermal amplification, and the reaction system of 25 mu L comprises: 12.5 μ L reaction buffer, 0.5 μ L dye, dyes including but not limited to color indicator and fluorescence indicator, 3.4 μ L primer and 3.4 μ L double distilled water. Then adding 5 mu L of double distilled water into the negative control of the control area as a negative control, and adding 5 mu L of BRAC1 gene into the positive control as a positive control; mu.L of the extracted RNA template was added to the detection zone. And sealing the micropore array chip added with the reaction solution by using a single-sided sealing film to form a sealed reaction chamber, and placing the micropore array chip on a hot plate at 60 ℃ to perform amplification reaction on the sample. The camera shooting component shoots color images or fluorescence images generated by the sample, and transmits the color images or fluorescence images to the main control computer for processing and analysis, and finally, the detection result is judged, and can be directly read by naked eyes in a fixed manner and can also be quantitatively displayed on the liquid crystal display screen.
The detection flux of the micropore array chip is adjusted according to the detection requirement, and 3-16 samples can be detected simultaneously. Meanwhile, a positive control and a negative control are added into the microporous array chip, the positive control adopts BRAC1 gene as the control, the influence of environmental factors on the amplification reaction is eliminated, the normal operation of the amplification reaction is ensured, and the false negative detection result which is easy to appear in the traditional nucleic acid detection process is avoided; the negative control adopts double distilled water as the control, so that the false positive result of constant-temperature amplification detection caused by environmental pollution is avoided, and the detection accuracy is improved.
For the primer design of the Loop-mediated isothermal amplification reaction, six primers are designed respectively aiming at the ORFlab/E/N/S gene region of the new coronavirus according to the requirement of the Loop-mediated isothermal amplification, wherein the six primers comprise an upstream inner primer (FIP), an upstream outer primer (F3), a downstream inner primer (BIP), a downstream outer primer (B3) and a Loop primer (LF and LB). And detecting the synthesized gene plasmids with different concentrations, wherein after the nucleic acid amplification reaction, the color of the positive sample is changed from pink to yellow, and the color of the negative sample is still pink.
The portable nucleic acid detector of the present invention can be applied to various commercially available kits, such as six respiratory virus nucleic acid detection kits (isothermal amplification chip method) of Chengdbo crystal core Biotechnology Co., Ltd.
The lowest detection limit of the nucleic acid detector is determined by two ways: the first method is to use a portable detector and a microwell array chip, and as can be seen from FIG. 5(A), a positive control in the control area reacts, while a negative control does not react, which proves that no false positive or false negative results appear during the detection process. As can be seen from the detection area, the detection sensitivity of the set of primers can reach 1000 copies/mL, and the sample volume is 5 muL; the second mode is real-time fluorescence detection, and as can be seen from fig. 5(B), on the surface of the real-time fluorescence curve, the reaction system can perform amplification detection on a detection target with a concentration of 1000 copies/mL or more, but no obvious reaction occurs on the detection target with a concentration of 100 copies/mL, so that the minimum detection limit is determined to be 1000 copies/mL. Thus proving that the portable nucleic acid detector and the micropore array chip can be effectively used for high-sensitivity detection of the new coronavirus.
Simultaneously, the same reaction solution and the detection target are respectively added into the micropore array chip and the commercial eight-connecting pipe, and are respectively placed on an electric hot plate. As can be seen in FIG. 6, the time required for detection of the microwell array chip was significantly lower than that of the commercial octal tube for the same detection concentration. The chip can realize amplification detection in 25min for different detection concentrations. This is because the chip is heated more uniformly on the hot plate, resulting in a faster reaction speed.
In one example, 130 clinical specimens were tested using the novel portable nucleic acid detector and microwell array chip of the present invention at Sichuan university Hospital, Washington, using throat swabs. All samples were tested by RT-qPCR and preliminary negative and positive results were obtained. Adopting a portable nucleic acid detector and a micropore array chip to detect the sample again, comparing the detection result with the RT-qPCR detection result, and if the PCR detection is positive and the chip detection is negative, judging the detection result as a false negative result; and if the PCR is negative and the chip is positive, the result is judged to be false positive. The detection accuracy of the system can reach 94.7 percent as found in the table 2. Meanwhile, the sensitivity and specificity of the clinical detection result are calculated as follows:
sensitivity ═ true positive results/(true positive results + false negative results);
specificity ═ true negative result/(true negative result + false positive result);
the sensitivity represents the capability of truly detecting a positive result, and the specificity represents the capability of truly detecting a negative result, and the calculation shows that the sensitivity of the primer system can reach 95.4 percent and the specificity reaches 95.35 percent, so that the designed portable nucleic acid detector has better detection performance on the new coronavirus.
Table 2.130 test results of clinical specimens
The micro-pore array chip and the portable detector for quickly detecting nucleic acid can be correspondingly adjusted according to different application scenes, and are designed and prepared in different ways. Therefore, the portable nucleic acid detector of the invention can be applied to large hospitals, and can be popularized to other application scenes, including:
1) a home detection scenario; the portable nucleic acid detector suitable for the scene can detect three parameters of ORF1ab gene, N gene and E gene of the new coronavirus of the same sample on the same chip; or the detection can be carried out aiming at the ORF1ab gene regions of three different samples, and the detection result is directly read by naked eyes, so that the cost is controlled and reduced as much as possible;
2) dense scenes of people streams such as airports and railway stations; the portable nucleic acid detector suitable for the scene adopts a high-flux micropore array chip, improves the detection flux, realizes rapid high-flux detection, and the rapid quantitative detection equipment is provided with a liquid crystal display screen and reads the results of a plurality of samples in a quantitative mode;
3) community hospital application scenarios: the portable nucleic acid detector suitable for the scene is provided with microporous detection chips with different detection fluxes according to the number of people in the community, and a quick reading device is provided with a liquid crystal display screen to read quantitative results and reduce the pressure of seeing a doctor of a large-scale medical institution;
4) overseas application scenarios: a high-cost quantitative reading all-in-one machine and a low-cost direct reading device are respectively provided for different countries and regions.
The foregoing is a more detailed description of the present patent with reference to specific preferred embodiments, and the specific embodiments of the patent are not to be considered limited to these descriptions. In addition, the related detection reagent can be suitable for the portable nucleic acid detector and other independently developed rapid detection systems, so that the field rapid detection of epidemic situations is realized.
The above-mentioned embodiments are intended to illustrate the objects, technical solutions and advantages of the present invention in further detail, and it should be understood that the above-mentioned embodiments are only exemplary embodiments of the present invention and are not intended to limit the present invention, and any modifications, equivalents, improvements and the like made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (10)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011642787.3A CN113150979A (en) | 2020-12-30 | 2020-12-30 | Portable nucleic acid detector based on micropore array chip |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202011642787.3A CN113150979A (en) | 2020-12-30 | 2020-12-30 | Portable nucleic acid detector based on micropore array chip |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113150979A true CN113150979A (en) | 2021-07-23 |
Family
ID=76878329
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202011642787.3A Pending CN113150979A (en) | 2020-12-30 | 2020-12-30 | Portable nucleic acid detector based on micropore array chip |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN113150979A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114854551A (en) * | 2022-04-27 | 2022-08-05 | 南京芳华基因科技有限公司 | Portable respiratory tract pathogenic bacteria nucleic acid detector |
| CN115109695A (en) * | 2022-06-20 | 2022-09-27 | 江苏猎阵生物科技有限公司 | Multifunctional PCR amplification equipment |
| CN115216395A (en) * | 2022-08-12 | 2022-10-21 | 西安交通大学 | A kind of automatic nucleic acid detection equipment and method |
| CN116083223A (en) * | 2023-02-28 | 2023-05-09 | 中国科学院微电子研究所 | A nucleic acid detection device and system |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP1447454A1 (en) * | 2003-02-14 | 2004-08-18 | DR. Chip Biotechnology Incorporation | Method and apparatus for detecting pathogens |
| CN1927466A (en) * | 2001-12-19 | 2007-03-14 | 阿菲梅特里克斯公司 | Array plate and manufacturing process for array plate assembly |
| CN102665916A (en) * | 2009-11-23 | 2012-09-12 | 3M创新有限公司 | Microwell array articles and methods of use |
| CN204714811U (en) * | 2015-02-09 | 2015-10-21 | 张国豪 | A kind of portable detection of nucleic acids equipment |
| WO2017025984A1 (en) * | 2015-08-07 | 2017-02-16 | Council Of Scientific And Industrial Research | Smartphone integrated real - time molecular diagnostic device |
| CN109536366A (en) * | 2018-11-29 | 2019-03-29 | 合肥中科易康达生物医学有限公司 | A kind of detection of nucleic acids micro-fluidic chip and nucleic acid detection system based on modified capillary |
| CN110564607A (en) * | 2019-09-11 | 2019-12-13 | 长春技特生物技术有限公司 | Full-automatic nucleic acid extraction amplification micro-fluidic chip dynamic quantitative detection integrated device |
-
2020
- 2020-12-30 CN CN202011642787.3A patent/CN113150979A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1927466A (en) * | 2001-12-19 | 2007-03-14 | 阿菲梅特里克斯公司 | Array plate and manufacturing process for array plate assembly |
| EP1447454A1 (en) * | 2003-02-14 | 2004-08-18 | DR. Chip Biotechnology Incorporation | Method and apparatus for detecting pathogens |
| CN102665916A (en) * | 2009-11-23 | 2012-09-12 | 3M创新有限公司 | Microwell array articles and methods of use |
| CN204714811U (en) * | 2015-02-09 | 2015-10-21 | 张国豪 | A kind of portable detection of nucleic acids equipment |
| WO2017025984A1 (en) * | 2015-08-07 | 2017-02-16 | Council Of Scientific And Industrial Research | Smartphone integrated real - time molecular diagnostic device |
| CN109536366A (en) * | 2018-11-29 | 2019-03-29 | 合肥中科易康达生物医学有限公司 | A kind of detection of nucleic acids micro-fluidic chip and nucleic acid detection system based on modified capillary |
| CN110564607A (en) * | 2019-09-11 | 2019-12-13 | 长春技特生物技术有限公司 | Full-automatic nucleic acid extraction amplification micro-fluidic chip dynamic quantitative detection integrated device |
Non-Patent Citations (2)
| Title |
|---|
| 动脉网: ""北航、华西医院联手：2周研发，5个月成型便捷式病原体核酸快速检测仪"", 《搜狐》 * |
| 北京航空航天大学常凌乾课题组: "北航、川大华西联合研发便携式病原体核酸快速检测仪", 《中国微米纳米技术学会》 * |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN114854551A (en) * | 2022-04-27 | 2022-08-05 | 南京芳华基因科技有限公司 | Portable respiratory tract pathogenic bacteria nucleic acid detector |
| CN115109695A (en) * | 2022-06-20 | 2022-09-27 | 江苏猎阵生物科技有限公司 | Multifunctional PCR amplification equipment |
| CN115216395A (en) * | 2022-08-12 | 2022-10-21 | 西安交通大学 | A kind of automatic nucleic acid detection equipment and method |
| CN116083223A (en) * | 2023-02-28 | 2023-05-09 | 中国科学院微电子研究所 | A nucleic acid detection device and system |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN113150979A (en) | Portable nucleic acid detector based on micropore array chip | |
| CN111337689A (en) | A novel coronavirus detection kit | |
| Tayyab et al. | Potential microfluidic devices for COVID-19 antibody detection at point-of-care (POC): A review | |
| US20240241059A1 (en) | Cartridge, system, and method for molecular diagnostic reaction testing | |
| JP2020042050A (en) | Test cartridge with integrated transfer module | |
| CN111077319A (en) | Micro-fluidic chip immunodetection kit and detection method thereof | |
| CN101614655B (en) | Detection method of microfluidic chip based on photoacoustic technique and device thereof | |
| CN112760193A (en) | Nucleic acid extraction/detection device and method | |
| CN104897907A (en) | Kit for detecting glycosylated hemoglobin and detection method thereof | |
| JP2006058280A (en) | Assay chip | |
| WO2020038235A1 (en) | High-flux biological, chemical and environmental detection system and method based on planar waveguide technology | |
| CN106442962A (en) | POCT instant detection device | |
| US20110053291A1 (en) | Method for analyzing sample solution and apparatus for analyzing sample solution | |
| CN117990897A (en) | Biochemical immunity all-in-one | |
| JP2016223919A (en) | Chip for chromatographic processing, chromatographic processing device, and chromatographic processing method | |
| Adedokun et al. | A handheld HIV detection platform using paper-based sample preparation and real-time isothermal amplification | |
| CN214750354U (en) | Novel coronavirus antigen and antibody combined intelligent detection device | |
| CN106198499A (en) | A kind of micro-fluidic chip for chemiluminescence detection and detection method thereof | |
| CN211339511U (en) | Integrated test strip for LAMP or RT-LAMP | |
| Liu et al. | A miniaturized platform based on reciprocating-flowing immunobinding for ultrafast, highly-sensitive and on-site immunoassay | |
| CN117778174A (en) | A microfluidic chip that simultaneously performs molecular diagnosis, immunodiagnosis, and biochemical detection and its testing method | |
| CN116338165A (en) | Protein chip kit for simultaneously detecting omazu and IgE | |
| CN112557684B (en) | Detection chip, detection equipment, detection system and detection method | |
| CN213398575U (en) | Novel test strip and kit for rapidly detecting coronavirus IgM/IgG | |
| JP7252925B2 (en) | Automated cloud-based detection system and method for detecting pathogens and antibody arrays at the point of care (POC) |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |