CN115612600A

CN115612600A - Automatic full-flow nucleic acid detection system and method

Info

Publication number: CN115612600A
Application number: CN202211242423.5A
Authority: CN
Inventors: 杨柯; 朱灵; 花昌义; 潘井宇; 王明龙; 朱灿灿; 赵俊; 邓国庆; 刘勇
Original assignee: Hefei Zhongke Yikangda Biomedical Co ltd
Current assignee: Hefei Zhongke Yikangda Biomedical Co ltd
Priority date: 2022-10-11
Filing date: 2022-10-11
Publication date: 2023-01-17

Abstract

The invention discloses an automatic full-flow nucleic acid detection system, wherein a microfluidic card box chip, a microfluidic control module, a temperature control module, a fluorescence detection module and a horizontal sliding guide rail are integrally arranged in a system bin; the microfluid control module is arranged above the temperature control module; the microfluidic cartridge chip can be mounted between the temperature control module and the microfluidic control module; the microfluid control module can move vertically up and down; the temperature control module and the fluorescence detection module are both arranged on the horizontal sliding guide rail. The invention also discloses a nucleic acid detection method. The system bin integrates a micro-fluid control module, a temperature control module and a fluorescence detection module, has the functions of nucleic acid extraction, amplification and detection, can perform automatic and totally-enclosed nucleic acid detection and analysis by combining a micro-fluid cartridge chip, realizes the whole automatic detection flow of 'sample inlet-result outlet', and has the advantages of high integration level, simple operation, pollution prevention, high sensitivity, low use technical requirement and the like.

Description

Automatic full-flow nucleic acid detection system and method

Technical Field

The invention relates to the technical field of medical instruments, in particular to an automatic full-flow nucleic acid detection system and method.

Background

Among the detection methods for SARS-CoV-2 (a novel coronavirus), nucleic acid detection technology (NAT) is gradually becoming the mainstream method for SARS-CoV-2 detection at present because of its advantages of high detection speed, high sensitivity and good specificity. A complete NAT process typically includes four steps: sample pretreatment, nucleic acid extraction, nucleic acid amplification and result detection; the whole process is complicated in steps and complex in operation, needs professional technicians to finish the process by using various matched instruments in a professional laboratory with complete matched facilities, and has the problem of false positive or false negative caused by the limitation of partial conditions or the error of the operators in the operation process. In addition, SARS-CoV-2 has high infectivity, and the complicated operation steps in the detection process can easily infect operators; therefore, the ideal nucleic acid detection method should be automated in a closed space against highly infectious pathogens such as SARS-CoV-2.

In the nucleic acid detection in the prior art, due to the limitation of an experimental method and experimental conditions, the operation process is complex and tedious, professional technicians and a professional laboratory are needed, the detection period is long, the risks of misoperation and infection of the laboratory personnel exist, and the pathogen detection can not be carried out in a resource-limited area. When dealing with a highly infectious pathogen such as SARS-CoV-2, the reactivity is insufficient. Therefore, an automatic full-flow nucleic acid detection system is urgently needed, the full-flow of nucleic acid extraction, amplification and detection can be automatically and totally carried out, professional technical personnel are not needed, pathogen detection in a resource-limited area is facilitated, meanwhile, the risk of infection of experimenters is reduced, and the detection sensitivity is improved.

The invention patent application with publication number CN113528326A discloses a microfluidic nucleic acid detection device and application. This application integrates the use of micro-fluidic nucleic acid detection chip through the cooperation, can accomplish to advance the automatic realization of back including nucleic acid extraction, amplification, full flow high flux nucleic acid detection and pathogen screening including the detection by nucleic acid testing equipment. However, the above-mentioned problems of complicated detection process and long detection period in nucleic acid detection still remain.

Disclosure of Invention

The technical problem to be solved by the invention is as follows: the problems of complicated detection process and long detection period of the existing nucleic acid detection are solved.

In order to solve the technical problems, the invention provides the following technical scheme:

an automatic full-flow nucleic acid detection system comprises a system bin, a microfluidic card box chip, a microfluidic control module, a temperature control module, a fluorescence detection module and a horizontal sliding guide rail;

the microfluidic card box chip, the microfluidic control module, the temperature control module, the fluorescence detection module and the horizontal sliding guide rail are all integrally installed in the system bin;

the microfluid control module is arranged above the temperature control module; the microfluidic cartridge chip can be mounted between the temperature control module and the microfluidic control module; the microfluidic control module can move vertically up and down;

the temperature control module and the fluorescence detection module are both arranged on the horizontal sliding guide rail, and the fluorescence detection module is connected with the temperature control module; during the moving process, the positions of the temperature control module and the fluorescence detection module are kept relatively fixed all the time.

The advantages are that: the system integrates the temperature control module, the microfluid control module and the fluorescence detection module in the system cabin, has the functions of nucleic acid extraction, amplification and detection, and has high system integration level; the automatic opening and closing of the system are guaranteed through the arrangement of the horizontal sliding guide rail, the chip is completely sealed in the whole experiment process, pollution is avoided, and the accuracy of an experiment result is improved.

Preferably, the temperature control module comprises a first heat sink, a second heat sink, a first temperature sensor, a second temperature sensor, a first peltier, a second peltier and a fin heat sink;

the first heat sink sheet is arranged at the upper end of the first Peltier, and the first temperature sensor is arranged inside the first heat sink sheet; the second heat sink sheet is arranged at the upper end of the second Peltier, and the second temperature sensor is arranged inside the second heat sink sheet;

the fin radiator is arranged in the middle of the temperature control module, the first Peltier is located between the first heat sink sheet and the fin radiator, and the second Peltier is located between the second heat sink sheet and the fin radiator.

Preferably, the temperature control module further comprises a fan; the fan is arranged at the bottom of the fin radiator.

Preferably, the microfluidic control module comprises a bracket, a stepping motor peristaltic pump, a direct current motor peristaltic pump, a two-position three-way electromagnetic valve, a lifting driving unit, a high-torque stepping motor, a photoelectric switch, an adapter, an air source connector base, a horizontal driving unit and a magnet;

the right side of the bracket is provided with an extending end, and the stepping motor peristaltic pump and the direct current motor peristaltic pump are fixedly arranged on the extending end; the two-position three-way electromagnetic valve is arranged on the right side of the bracket;

the high-torque stepping motor, the photoelectric switch, the conversion piece and the air source connector base are vertically and slidably arranged at the front end of the rear part of the bracket through the lifting driving unit;

the photoelectric switch, the conversion piece and the air source connector base are all fixedly arranged at the bottom of the lifting driving unit;

the step motor peristaltic pump and the direct current motor peristaltic pump are respectively connected with a two-position three-way electromagnetic valve through air ducts, and the two-position three-way electromagnetic valve is connected with an air inlet on the microfluidic card box chip through the air duct and an air source connector seat; the high-torque stepping motor is connected with the microfluidic card box chip through the adapter;

the horizontal driving unit is arranged at the bottom of the bracket in a sliding manner, the magnet is arranged on the horizontal driving unit, and the magnet is positioned on the side surface close to the nucleic acid extraction tube on the microfluidic card box chip.

Preferably, two collision switches are provided at both ends of the horizontal driving unit.

Preferably, the fluorescence detection module comprises a support plate, a turntable, an LED lamp bead, an excitation filter, an excitation focusing lens, an optical fiber, an emission focusing lens group, an emission filter and a detector;

the turntable is rotatably arranged on the supporting plate;

the LED lamp beads are fixedly arranged on the rotary table, and the excitation optical filter is arranged at the front ends of the LED lamp beads; the detector is fixed on the supporting plate, and the emission optical filter is placed at the front end of the detector; the excitation filter and the emission filter are both fixed on the turntable;

the excitation focusing lens and the emission focusing lens group are fixed on the support plate; the excitation focusing lens is arranged at the front end of the excitation optical filter, and the emission focusing lens group is arranged at the front end of the emission optical filter;

one end of the optical fiber is fixed on the side surface of the second heat sink sheet of the temperature control module, and the other end of the optical fiber is fixed on the supporting plate and is respectively arranged at the front ends of the excitation focusing lens and the shooting focusing lens group.

Preferably, the optical fiber is a Y-shaped glass optical fiber, a beam combining end of the optical fiber is fixed on a side surface of the second heat sink sheet of the temperature control module, and two beam splitting ends are fixed on the supporting plate and respectively connected with the LED lamp bead and the detector correspondingly;

the excitation focusing lens is arranged behind the optical fiber beam splitting end corresponding to the LED lamp bead, and the emission focusing lens group is arranged behind the optical fiber beam splitting end corresponding to the detector.

Preferably, the LED lamp bead is provided with a plurality of different wavelengths, and the excitation optical filter and the emission optical filter are respectively and correspondingly provided with a plurality of groups.

Preferably, the fluorescence detection module further comprises a stepping drive motor, and an output shaft of the stepping drive motor is fixed at the center of the turntable.

The invention also discloses a method for adopting the automatic full-flow nucleic acid detection system, which comprises the following steps:

s1, loading a collected sample into a microfluidic card box chip;

s2, clicking a 'bin opening' button, driving the temperature control module and the fluorescence detection module to move by the horizontal sliding guide rail, and automatically opening the bin; placing the microfluidic card box chip subjected to sample loading in a limiting groove on the upper surface of a temperature control module, wherein a sample cracking tube and a nucleic acid amplification tube at the bottom of the chip are respectively tightly attached to a corresponding first heat sink sheet and a corresponding second heat sink sheet; clicking a 'closing cabin' button, driving the temperature control module and the fluorescence detection module to move by the horizontal sliding guide rail, and automatically closing the cabin to finish the chip on-machine operation;

s3, clicking a start button, firstly, calibrating zero of each moving part in the system, then, moving the lifting driving unit downwards, and butting the adaptor and the air source connector seat with the microfluidic card box chip;

s4, the temperature of the first Peltier on the temperature control module starts to rise, the first temperature sensor detects the temperature of the first heat sink piece in real time, and when the temperature reaches a set temperature, the temperature is kept for a period of time at a constant temperature according to set time;

s5, after the sample is cracked, according to a preset nucleic acid extraction process, starting a stepping motor peristaltic pump, a direct current motor peristaltic pump, a two-position three-way electromagnetic valve, a high-torque stepping motor, a photoelectric switch, a linear sliding guide rail and a collision switch, and automatically completing the steps of nucleic acid adsorption, cleaning, elution, liquid separation and sealing;

s6, after the nucleic acid extraction is finished, the temperature of a second Peltier on the temperature control module begins to rise, a second temperature sensor detects the temperature of a second heat sink sheet in real time, accurate temperature control is carried out according to a preset temperature condition, and meanwhile, a fluorescence acquisition module carries out fluorescence data acquisition according to the preset condition;

and S7, after the nucleic acid amplification is finished, automatically correcting zero of each moving part in the system, and analyzing the acquired fluorescence data to give an experimental result.

Compared with the prior art, the invention has the beneficial effects that:

(1) The automatic full-flow nucleic acid detection system integrates a temperature control module, a microfluid control module and a fluorescence detection module, has the functions of nucleic acid extraction, amplification and detection, and has high system integration level.

(2) The automatic full-flow nucleic acid detection system provided by the invention is combined with the microfluidic card box chip, the full flow from nucleic acid extraction, amplification and detection can be automatically carried out on the chip, the experiment operation is simple, and the automation degree is high.

(3) When the automatic full-flow nucleic acid detection system is used in a laboratory, the interior of the chip is completely sealed in the whole experiment process, so that pollution is avoided, the risk of infection of experimenters is reduced, and the accuracy of experiment results is improved.

(4) The fluorescence detection module of the automatic full-flow nucleic acid detection system integrates various fluorescence detection wave bands, can detect various pathogens in one detection tube, and has a multi-target detection function.

(5) The automatic full-flow nucleic acid detection system is simple to operate, low in technical threshold, free of professional experiment skills for operators and convenient to popularize.

Drawings

FIG. 1 is a schematic diagram of the overall structure of an automated full-flow nucleic acid detection system according to an embodiment of the present invention;

FIG. 2 is a schematic diagram of a temperature control module of an automated full-flow nucleic acid detection system according to an embodiment of the present invention;

FIG. 3 is a schematic diagram of a microfluidic control module of an automated full-flow nucleic acid detection system according to an embodiment of the present invention;

FIG. 4 is a schematic diagram of a fluorescence detection module of an automated full-flow nucleic acid detection system according to an embodiment of the present invention;

FIG. 5 is a front view of a fluorescence detection module of an automated full-flow nucleic acid detection system according to an embodiment of the present invention;

in the figure: 1. a microfluidic cartridge chip; 2. a temperature control module; 21. a first heat sink sheet; 22. a second heat sink sheet; 23. a first temperature sensor; 24. a second temperature sensor; 25. a first peltier; 26. A second peltier; 27. a fin radiator; 28. a fan; 3. a microfluidic control module; 31. a support; 32. a stepper motor peristaltic pump; 33. a direct current motor peristaltic pump; 34. a two-position three-way electromagnetic valve; 35. a lifting drive unit; 36. a high torque stepper motor; 37. a photoelectric switch; 38. an adapter; 39. an air source joint seat; 310. a horizontal driving unit; 311. a magnet; 312. a bump switch; 4. a fluorescence detection module; 41. a support plate; 42. a turntable; 43. a step-by-step drive motor; 44. LED lamp beads; 45. exciting the optical filter; 46. exciting the focusing lens; 47. an optical fiber; 48. an emission focusing lens group; 49. an emission filter; 410. a detector; 5. a horizontal sliding guide rail.

Detailed Description

In order to facilitate the understanding of the technical solutions of the present invention for those skilled in the art, the technical solutions of the present invention will be further described with reference to the drawings attached to the specification.

The terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present application, "a plurality" means two or more unless specifically limited otherwise.

Referring to fig. 1, the present embodiment discloses an automated full-flow nucleic acid detection system, which includes a system chamber (not shown), a microfluidic cartridge chip 1, a temperature control module 2, a microfluidic control module 3, a fluorescence detection module 4, and a horizontal sliding guide rail 5. The temperature control module 2, the microfluid control module 3, the fluorescence detection module 4 and the controller module are all integrated in the accommodating bin. Through with each module integration in inclosed system storehouse, guarantee whole implementation process chip inside and seal completely, avoid polluting, reduced the risk that the experimenter infects, also improved the accuracy of experimental result.

The temperature control module 2 is positioned at the bin inlet of the system, and a limiting groove for placing the microfluidic card box chip 1 is arranged above the temperature control module. The microfluidic control module 3 is located above the temperature control module 2 and can move in the vertical direction to facilitate automated docking with the microfluidic cartridge chip 1. Fluorescence detection module 4 is located temperature control module 2 rear, and both install on horizontal sliding guide 5, and accessible step motor moves in the horizontal direction, and both positions remain relatively fixed throughout at removal in-process, the automatic storehouse of opening and closing of system of being convenient for.

Referring to fig. 2, the temperature control module 2 includes a first heat sink 21, a second heat sink 22, a first temperature sensor 23, a second temperature sensor 24, a first peltier 25, a second peltier 26, a fin radiator 27, and a fan 28.

The first heat sink 21 is arranged at the upper end of the first Peltier 25, and the first temperature sensor 23 is arranged inside the first heat sink 21; the second heat sink 22 is mounted on the upper end of the second peltier element 26, and the second temperature sensor 24 is mounted inside the second heat sink 22. The first heat sink piece 21 is used for providing uniform temperature conditions for sample cracking on the microfluidic cartridge chip 1, and the second heat sink piece 22 is used for providing uniform temperature conditions for nucleic acid amplification on the microfluidic cartridge chip 1; the first temperature sensor 23 and the second temperature sensor 24 detect the temperatures of the first heat sink 21 and the second heat sink 22, respectively, in real time.

The fin radiator 27 is disposed at the middle of the temperature control module 2, the first peltier element 25 is located between the first heat sink sheet 21 and the fin radiator 27, and the second peltier element 26 is located between the second heat sink sheet 22 and the fin radiator 27. The first peltier 25 is used for real-time temperature control according to the set temperature and the detected temperature of the first temperature sensor 23; the second peltier 26 is used for real-time temperature control according to the set temperature and the detected temperature of the second temperature sensor 24, respectively. The fan 28 is located at the bottom of the fin radiator 27 and is used together with the fin radiator 27 for module heat dissipation.

The first heat sink 21, the first temperature sensor 23 and the first peltier 25 are used for sample lysis on the microfluidic cartridge chip 1, and the first heat sink 21 is a single well. The second heat sink 22, the second temperature sensor 24 and the second peltier 26 are used for nucleic acid amplification on the microfluidic cartridge chip 1, and the number of the wells of the second heat sink 22 varies with the number of the detection tubes on the microfluidic cartridge chip 1.

Referring to fig. 3, the microfluidic control module 3 includes a bracket 31, a step motor peristaltic pump 32, a dc motor peristaltic pump 33, a two-position three-way solenoid valve 34, a lifting driving unit 35, a high torque step motor 36, an electro-optical switch 37, an adaptor 38, an air source connector base 39, a horizontal driving unit 310, a magnet 311, and a collision switch 312.

The right side of the bracket 31 is provided with an extending end, and the stepping motor peristaltic pump 32 and the direct current motor peristaltic pump 33 are fixedly arranged on the extending end; a two-position three-way solenoid valve 34 is installed on the right side of the bracket 31.

The high-torque stepping motor 36, the photoelectric switch 37, the converter and the air source connector base 39 are vertically and slidably mounted at the front end of the rear part of the bracket 31 through the lifting driving unit 35; a high torque stepper motor 36 is connected to the elevation drive unit 35 for powering the elevation drive unit 35. The photoelectric switch 37, the conversion piece and the air source connector base 39 are all fixedly arranged at the bottom of the lifting driving unit 35.

The step motor peristaltic pump 32 and the direct current motor peristaltic pump 33 are respectively connected with a two-position three-way electromagnetic valve 34 through air ducts, and the two-position three-way electromagnetic valve 34 is connected with an air inlet on the microfluidic cartridge chip 1 through the air duct and an air source connector seat 39. The stepper motor peristaltic pump 32 allows for precise liquid transfer in the microfluidic cartridge chip 1 by controlling the rotation angle of the stepper motor, and the dc motor peristaltic pump 33 allows for sufficient liquid mixing in the microfluidic cartridge chip 1 by high speed rotation of the dc motor. By controlling the conduction mode of the two-position three-way electromagnetic valve 34, the step motor peristaltic pump 32 or the direct current motor peristaltic pump 33 can be respectively controlled to be connected into the microfluidic cartridge chip 1 to perform different microfluidic operations.

The high-torque stepping motor 36 is connected with the microfluidic cartridge chip 1 through the adapter piece 38, the photoelectric switch 37 calibrates the initial position of the high-torque stepping motor 36, and the micro-channel switching of the microfluidic cartridge chip 1 can be accurately performed by controlling the rotation angle of the motor. The lifting driving unit 35 can be controlled to move in the vertical direction through the rotation of the high-torque stepping motor 36, so that the automatic butt joint and separation of the adaptor 38 and the air source connector base 39 and the microfluidic cartridge chip 1 are realized.

The horizontal driving unit 310 is slidably mounted on the bottom of the rack 31, the magnet 311 is mounted on the horizontal driving unit 310, and the magnet 311 is located near the side of the nucleic acid extraction tube on the microfluidic cartridge chip 1. The magnet 311 is used for adsorbing magnetic beads in the nucleic acid extracted by the magnetic bead method. When the magnetic bead method is used for extracting nucleic acid, the magnet 311 can move in the horizontal direction by the movement of the horizontal driving unit 310, and sequentially passes through the side surfaces of the nucleic acid extraction tube on the microfluidic cartridge chip 1, so that when the magnet 311 approaches the extraction tube, the magnetic beads in the tube are adsorbed, and when the magnet 311 leaves the extraction tube, the magnetic beads in the tube are desorbed.

Meanwhile, two collision switches 312 are provided at both ends of the horizontal driving unit 310 for limiting the position of the magnet 311.

Referring to fig. 4 and 5, the fluorescence detection module 4 includes a support plate 41, a turntable 42, a stepping motor 43, an LED lamp bead 44, an excitation filter 45, an excitation focusing lens 46, an optical fiber 47, an emission focusing lens set 48, an emission filter 49, and a detector 410.

The LED lamp beads 44 are fixedly arranged on the rotary table 42, and the excitation light filter 45 is arranged at the front ends of the LED lamp beads 44; the detector 410 is fixed to the support plate 41, and the emission filter 49 is placed at the front end of the detector 410.

Optical fiber 47 is Y type glass optical fiber 47, and the end of restrainting of combining of optical fiber 47 is fixed in the side of 2 second heat sink pieces 22 of temperature control module, and two beam splitting ends are all fixed on backup pad 41, and correspond with LED lamp pearl 44 and detector 410 respectively and are connected.

Meanwhile, the excitation focusing lens 46 is arranged behind the beam splitting end of the optical fiber 47 corresponding to the LED lamp bead 44, and the emission focusing lens group 48 is arranged behind the beam splitting end of the optical fiber 47 corresponding to the detector 410.

The excitation focus lens 46 and the emission focus lens group 48 are fixed to the support plate 41. Excitation light filter 45 and emission light filter 49 are all fixed on carousel 42, and step drive motor 43's output shaft is fixed in the center of carousel 42, drives carousel 42 rotatory through controlling step drive motor 43, can drive LED lamp pearl 44, excitation light filter 45 and emission light filter 49 rotation in step.

Excitation light emitted by the LED lamp bead 44 enters a beam splitting end of the optical fiber 47 through the filtering of the excitation filter 45 and the focusing of the excitation focusing lens 46, and is emitted to a detection tube of the microfluidic card box chip 1 at a beam combining end of the optical fiber 47 to excite fluorescence. Part of the fluorescence is transmitted through the optical fiber 47, exits at the other beam splitting end, passes through the focusing of the emission focusing lens group 48 and the filtering of the emission filter 49, and is received by the detector 410.

Meanwhile, the LED lamp beads 44 can be set to have a plurality of different wavelengths, and the excitation filter 45 and the emission filter 49 are set to have corresponding wavelengths so as to realize fluorescence detection of different wave bands. In this embodiment, six sets of LED lamp beads 44 and corresponding optical filters are provided, so that fluorescence detection of six different wavelength bands, i.e., FAM, HEX, TAMRA, ROX, CY5, and CY5.5, can be achieved. The number of the optical fibers 47 and the number of the detectors 410 are the same and correspond to the number of the detection tubes on the microfluidic cartridge chip 1.

In this embodiment, the stepping driving motor 43 is controlled to drive the turntable 42 to rotate, so as to switch the LED lamp beads 44, the excitation filter 45 and the emission filter 49, and realize the fluorescence detection of various different wave bands in one detection tube.

The automatic full-flow nucleic acid detection system of the embodiment integrates the temperature control module 2, the micro-fluid control module 3 and the fluorescence detection module 4, has the functions of nucleic acid extraction, amplification and detection, and has high system integration level.

The automatic full-flow nucleic acid detection system of the embodiment combines the microfluidic card box chip 1, can automatically perform the full flow from nucleic acid extraction to amplification to detection on the chip, and has simple experiment operation and high automation degree.

The automatic full-flow nucleic acid detection system of this embodiment is when carrying out the laboratory, and whole experimentation chip is inside to be sealed totally, avoids polluting, has reduced the risk that the experimenter infects promptly, has also improved the accuracy of experimental result.

The fluorescence detection module 4 of the automatic full-flow nucleic acid detection system of the embodiment integrates various fluorescence detection wave bands, can detect various pathogens in one detection tube, and has a multi-target detection function.

The automatic full-flow nucleic acid detection system of this embodiment easy operation, the technical threshold is low, and operating personnel need not to possess professional experimental skill, the facilitate promotion.

The automatic nucleic acid detection full-flow steps of the detection system are as follows:

(1) Loading the collected sample into the microfluidic cartridge chip 1;

(2) Clicking the button of opening the warehouse, driving the temperature control module 2 and the fluorescence detection module 4 to move by the horizontal sliding guide rail 5, and automatically opening the warehouse. The microfluidic cartridge chip 1 which finishes sample loading is placed in a limiting groove on the upper surface of the temperature control module 2, and a sample cracking tube and a nucleic acid amplification tube at the bottom of the chip are respectively clung to the corresponding first heat sink sheet 21 and the second heat sink sheet 22. And clicking a 'closing cabin' button, driving the temperature control module 2 and the fluorescence detection module 4 to move by the horizontal sliding guide rail 5, automatically closing the cabin, and finishing the operation of the chip on the computer.

(3) Clicking the "start" button first performs zeroing of the various moving parts inside the system, and then the lift drive unit 35 moves down to perform docking of the adapter piece 38 and the gas source connector holder 39 with the microfluidic cartridge chip 1.

(4) The temperature of the first peltier 25 on the temperature control module 2 starts to rise, the first temperature sensor 23 detects the temperature of the first heat sink 21 in real time, and when the temperature reaches the set temperature, the temperature is kept for a period of time at a constant temperature according to the set time.

(5) After the sample is cracked, according to a preset nucleic acid extraction process, the step motor peristaltic pump 32, the direct current motor peristaltic pump 33, the two-position three-way electromagnetic valve 34, the high-torque step motor 36, the photoelectric switch 37, the linear sliding guide rail and the collision switch 312 are started, and steps of nucleic acid adsorption, cleaning, elution, liquid separation and sealing are automatically completed.

(6) After the nucleic acid extraction is completed, the temperature of the second peltier 26 on the temperature control module 2 starts to rise, the second temperature sensor 24 detects the temperature of the second heat sink 22 in real time, the temperature is accurately controlled according to the preset temperature conditions, and meanwhile, the fluorescence data acquisition module acquires fluorescence data according to the preset conditions.

(7) After the nucleic acid amplification is finished, automatically zeroing each moving part in the system, and simultaneously analyzing the collected fluorescence data to give an experimental result.

It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein, and any reference signs in the claims are not intended to be construed as limiting the claim concerned.

The above-mentioned embodiments only represent embodiments of the present invention, and the protection scope of the present invention is not limited to the above-mentioned embodiments, and it will be apparent to those skilled in the art that several variations and modifications can be made without departing from the concept of the present invention, and these embodiments are all within the protection scope of the present invention.

Claims

1. An automated full-flow nucleic acid detection system, comprising: comprises a system bin, a microfluidic card box chip (1), a microfluidic control module (3), a temperature control module (2), a fluorescence detection module (4) and a horizontal sliding guide rail (5);

the microfluidic card box chip (1), the microfluidic control module (3), the temperature control module (2), the fluorescence detection module (4) and the horizontal sliding guide rail (5) are integrally installed in the system bin;

the microfluid control module (3) is arranged above the temperature control module (2); the microfluidic cartridge chip (1) can be installed between the temperature control module (2) and the microfluidic control module (3); the microfluid control module (3) can move vertically up and down;

the temperature control module (2) and the fluorescence detection module (4) are both arranged on the horizontal sliding guide rail (5), and the fluorescence detection module (4) is connected with the temperature control module (2); during the moving process, the positions of the temperature control module (2) and the fluorescence detection module (4) are always kept relatively fixed.

2. The automated full-flow nucleic acid detection system according to claim 1, wherein: the temperature control module (2) comprises a first heat sink sheet (21), a second heat sink sheet (22), a first temperature sensor (23), a second temperature sensor (24), a first Peltier (25), a second Peltier (26) and a fin radiator (27);

the first heat sink piece (21) is arranged at the upper end of the first Peltier (25), and the first temperature sensor (23) is arranged inside the first heat sink piece (21); the second heat sink sheet (22) is arranged at the upper end of the second Peltier (26), and the second temperature sensor (24) is arranged inside the second heat sink sheet (22);

the fin radiator (27) is arranged in the middle of the temperature control module (2), the first Peltier (25) is positioned between the first heat sink sheet (21) and the fin radiator (27), and the second Peltier (26) is positioned between the second heat sink sheet (22) and the fin radiator (27).

3. The automated full-flow nucleic acid detection system according to claim 2, wherein: the temperature control module (2) further comprises a fan (28); the fan (28) is mounted at the bottom of the fin heat sink (27).

4. The automated full-flow nucleic acid detection system according to claim 1, wherein: the micro-fluid control module (3) comprises a bracket (31), a stepping motor peristaltic pump (32), a direct current motor peristaltic pump (33), a two-position three-way electromagnetic valve (34), a lifting driving unit (35), a high-torque stepping motor (36), a photoelectric switch (37), an adapter (38), an air source connector base (39), a horizontal driving unit (310) and a magnet (311);

an extending end is arranged on the right side of the bracket (31), and the stepping motor peristaltic pump (32) and the direct current motor peristaltic pump (33) are fixedly arranged on the extending end; the two-position three-way electromagnetic valve (34) is arranged on the right side of the bracket (31);

the high-torque stepping motor (36), the photoelectric switch (37), the conversion piece and the air source connector base (39) are vertically and slidably arranged at the front end of the rear part of the bracket (31) through the lifting driving unit (35);

the photoelectric switch (37), the conversion piece and the air source connector base (39) are fixedly arranged at the bottom of the lifting driving unit (35);

the step motor peristaltic pump (32) and the direct current motor peristaltic pump (33) are respectively connected with a two-position three-way electromagnetic valve (34) through air ducts, and the two-position three-way electromagnetic valve (34) is connected with an air inlet on the microfluidic card box chip (1) through the air duct and an air source connector seat (39); the high-torque stepping motor (36) is connected with the microfluidic card box chip (1) through an adapter piece (38);

the horizontal driving unit (310) is arranged at the bottom of the bracket (31) in a sliding way, the magnet (311) is arranged on the horizontal driving unit (310), and the magnet (311) is positioned close to the side surface of the nucleic acid extraction tube on the microfluidic cartridge chip (1).

5. The automated full-flow nucleic acid detection system according to claim 4, wherein: two collision switches (312) are arranged at two ends of the horizontal driving unit (310).

6. The automated full-flow nucleic acid detection system of claim 1, wherein: the fluorescence detection module (4) comprises a supporting plate (41), a rotary table (42), LED lamp beads (44), an excitation filter (45), an excitation focusing lens (46), an optical fiber (47), an emission focusing lens group (48), an emission filter (49) and a detector (410);

the rotary disc (42) is rotatably arranged on the supporting plate (41);

the LED lamp beads (44) are fixedly arranged on the rotary table (42), and the excitation optical filter (45) is arranged at the front ends of the LED lamp beads (44); the detector (410) is fixed on the supporting plate (41), and the emission filter (49) is placed at the front end of the detector (410); the excitation filter (45) and the emission filter (49) are both fixed on the turntable (42);

the excitation focusing lens (46) and the emission focusing lens group (48) are fixed on the support plate (41); the excitation focusing lens (46) is arranged at the front end of the excitation filter (45), and the emission focusing lens group (48) is arranged at the front end of the emission filter (49);

one end of the optical fiber (47) is fixed on the side surface of the second heat sink sheet (22) of the temperature control module (2), and the other end of the optical fiber is fixed on the supporting plate (41) and is respectively arranged at the front ends of the excitation focusing lens (46) and the shooting focusing lens group.

7. The automated full-flow nucleic acid detection system according to claim 6, wherein: the optical fiber (47) is a Y-shaped glass optical fiber (47), the beam combining end of the optical fiber (47) is fixed on the side surface of the second heat sink sheet (22) of the temperature control module (2), and the two beam splitting ends are fixed on the supporting plate (41) and are respectively correspondingly connected with the LED lamp bead (44) and the detector (410);

the excitation focusing lens (46) is placed behind the optical fiber (47) beam splitting end corresponding to the LED lamp bead (44), and the emission focusing lens group (48) is placed behind the optical fiber (47) beam splitting end corresponding to the detector (410).

8. The automated full-flow nucleic acid detection system according to claim 6, wherein: the LED lamp beads (44) are provided with a plurality of different wavelengths, and the excitation filter (45) and the emission filter (49) are respectively and correspondingly provided with a plurality of groups.

9. The automated full-flow nucleic acid detection system according to claim 6, wherein: the fluorescence detection module (4) further comprises a stepping drive motor (43), and an output shaft of the stepping drive motor (43) is fixed at the center of the turntable (42).

10. A method of using the automated full-flow nucleic acid detection system of any one of claims 1-9, wherein: the method comprises the following steps:

s1, loading the collected sample into a microfluidic card box chip (1);

s2, clicking a 'bin opening' button, driving the temperature control module (2) and the fluorescence detection module (4) to move by the horizontal sliding guide rail (5), and automatically opening the bin; placing the microfluidic cartridge chip (1) which finishes sample loading in a limiting groove on the upper surface of a temperature control module (2), wherein a sample cracking tube and a nucleic acid amplification tube at the bottom of the chip are respectively clung to a corresponding first heat sink sheet (21) and a corresponding second heat sink sheet (22); clicking a 'closing cabin' button, driving the temperature control module (2) and the fluorescence detection module (4) to move by the horizontal sliding guide rail (5), and automatically closing the cabin to finish the operation of the chip on the computer;

s3, clicking a start button, firstly, calibrating zero of each moving part in the system, then moving the lifting driving unit (35) downwards, and butting the adaptor (38) and the air source connector seat (39) with the microfluidic cartridge chip (1);

s4, a first Peltier (25) on the temperature control module (2) starts to heat up, a first temperature sensor (23) detects the temperature of a first heat sink piece (21) in real time, and after the temperature reaches a set temperature, the temperature is kept for a period of time at a constant temperature according to set time;

s5, after the sample is cracked, according to a preset nucleic acid extraction flow, starting a stepping motor peristaltic pump (32), a direct current motor peristaltic pump (33), a two-position three-way electromagnetic valve (34), a high-torque stepping motor (36), a photoelectric switch (37), a linear sliding guide rail and a collision switch (312), and automatically completing steps of nucleic acid adsorption, cleaning, elution, liquid separation and sealing;

s6, after nucleic acid extraction is finished, the temperature of a second Peltier (26) on the temperature control module (2) begins to rise, a second temperature sensor (24) detects the temperature of a second heat sink sheet (22) in real time, accurate temperature control is carried out according to preset temperature conditions, and meanwhile, a fluorescence acquisition module carries out fluorescence data acquisition according to the preset conditions;

and S7, after the nucleic acid amplification is finished, automatically zeroing each moving part in the system, and analyzing the collected fluorescence data to give an experimental result.