Full-automatic PCR (Polymerase Chain Reaction) analyzer based on micro-fluidic card box and nucleic acid detection method

Abstract

According to the full-automatic PCR analyzer based on the microfluidic card box, a fixing frame is fixedly connected into an installation shell, an installation plate is fixed to the fixing frame, and an inserting-connecting sinking groove is formed in the side, opposite to a door body, of the installation plate; the lower end of the reagent reaction box can be inserted into the mounting shell through the inserting sinking groove, the reagent reaction box comprises a bottom plate, the upper side of the bottom plate is fixedly connected with a reagent box body, and the reagent box body is provided with a channel communicating control hole; a plurality of reagent liquid storage tanks and amplification liquid tanks are arranged on the upper side of the kit body outside one end of the channel communicated control hole in the left-right direction; the nucleic acid extraction assembly is arranged in the mounting shell, the nucleic acid extraction assembly comprises a stirring magnetic rod which can ascend and descend and can move in the front-back direction and the left-right direction, the stirring magnetic rod can stretch into any reagent liquid storage pool, and a magnetic separation module which can ascend and descend is connected to the portion, below the mounting plate, of the fixing frame; the nucleic acid extraction device can realize automatic extraction of nucleic acid and is convenient to amplify.

Description

Technical Field

[0001] The invention relates to the field of microfluidic technology, in particular to a full-automatic PCR analyzer based on a microfluidic cartridge. Background Art

[0002] Since its invention, PCR technology has been widely recognized and applied in the life science community. It is not only used in the medical field to detect and diagnose infectious diseases, genetic diseases, tumors and other diseases, but also plays a pivotal role in biological research, promoting the development of genetic engineering, gene sequencing, gene mutation analysis and other fields. In addition, PCR technology also has important applications in forensic medicine and environmental science, such as DNA identification and matching of criminal crimes, detection and monitoring of environmental pollutants, etc.

[0003] The principle of fluorescence quantitative PCR is to add fluorescent groups to the PCR reaction system, which accumulate continuously as the DNA is amplified. By detecting the changes in the fluorescence signal in real time, the changes in the amount of the product during the PCR amplification process can be accurately reflected. At the same time, the starting template can be quantitatively analyzed using the analysis of the Ct value and the standard curve. In the field of basic scientific research, fluorescence quantitative PCR is widely used in gene expression analysis and abundance detection. By detecting the fluorescence signal of a specific gene in cells, tissues or samples, the expression level of the gene and its distribution in the organism can be accurately obtained, which is of great significance for understanding gene function, revealing the laws of growth and development of organisms, and exploring the mechanism of disease occurrence.

[0004] For fluorescent quantitative PCR, people have found that by adding fluorescent groups and quenching groups to TaqMan probes for amplification, the fluorescent groups are released after each amplification, and large-scale samples can be quantitatively analyzed in real time. At the same time, by replacing the fluorescent groups on the TaqMan probes used for amplification of different nucleic acids, real-time analysis and detection of multiple pathogens can be achieved simultaneously. This technology has also been widely used in the field of PCR. In the existing technology, nucleic acids in organisms are artificially extracted, and the extraction rate of nucleic acids is easily affected by temperature, humidity and human factors. When analyzing large-scale samples, human factors and environmental factors will affect the final PCR analysis; in addition, when performing multi-pathogen detection, fluorescent groups with similar wavelengths will not be used for fluorescent quantitative PCR analysis in the same system, which is not suitable for the complexity trend of pathogen detection kits currently on the market. Summary of the invention

[0005] The purpose of this section is to summarize some aspects of embodiments of the present invention and briefly introduce some preferred embodiments. Some simplifications or omissions may be made in this section and the specification abstract and the invention title of this application to avoid blurring the purpose of this section, the specification abstract and the invention title, and such simplifications or omissions cannot be used to limit the scope of the present invention.

[0006] In view of the above and / or existing problems in the existing PCR analysis, the present invention is proposed.

[0007] Therefore, the purpose of the present invention is to provide a fully automatic PCR analyzer based on a microfluidic cartridge, which solves the technical problems that the extraction rate of nucleic acid extracted from organisms is unstable, affecting the amplification effect of nucleic acid and thus affecting the detection sensitivity, and fluorescent groups with similar wavelengths cannot be used for fluorescent quantitative PCR analysis in the same system, thereby reducing the multiplicity of PCR analysis. The present invention can realize the automated extraction of nucleic acid and facilitate amplification.

[0008] In order to solve the above technical problems, the present invention provides the following technical solutions: a fully automatic PCR analyzer based on a microfluidic cartridge, comprising: A mounting shell having a receiving cavity, wherein one side of the mounting shell in the left-right direction is connected to a door body that can be opened and closed, a fixed frame is fixedly connected inside the mounting shell, a mounting plate is fixed on the fixed frame, and a plug-in sink groove is opened on one side of the mounting plate opposite to the door body; A reagent reaction box, wherein the lower end of the reagent reaction box can be inserted into the mounting shell through the plug-in sink, the reagent box comprises a bottom plate, the upper side of the bottom plate is fixedly connected with a reagent box body, a channel connection control hole is opened on the reagent box body, the upper side of the reagent box body outside one end of the channel connection control hole in the left and right directions has a plurality of reagent storage pools and amplification liquid pools, the upper part of the reagent box body at the other end of the channel connection control hole in the left and right directions has a plurality of probe pools that can be connected with the amplification liquid pools, and the probe pool has an amplification chamber on the reagent box body at one end of the left and right directions away from the channel connection hole; A nucleic acid extraction component is arranged in the installation shell, and the nucleic acid extraction component includes a stirring magnetic bar that can be raised and lowered and can move in the front-back direction and the left-right direction. The stirring magnetic bar can extend into any reagent storage tank. A magnetic separation module that can be raised and lowered is connected to the fixed frame below the installation plate.

[0009] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, wherein: the reagent box body is just movably connected with a control communication valve body through a channel communication control hole, and a plurality of reagent storage tanks are respectively a first reagent storage tank, a second reagent storage tank, a third reagent storage tank and a fourth reagent storage tank, and a drainage hole is opened at the bottom of the fourth reagent storage tank, and a first channel is opened on the reagent box body below the drainage hole pool, and an inlet and outlet hole is opened at the bottom of the amplification liquid pool, and a second channel is opened on the reagent box body below the inlet and outlet hole, and the other end of the first channel can be connected to the amplification liquid pool through the second channel and the inlet and outlet hole in sequence; in the present invention, the magnetic separation module includes an electromagnetic separation seat, and the upper end of the electromagnetic separation seat is opened with a fourth reagent The liquid storage tanks correspond to magnetic separation ports one by one, and an electromagnet is connected to the electromagnetic separation seat at the bottom of the magnetic separation port (which is the prior art). A support part for facilitating the flow of liquid is fixed on the reagent box body at the bottom of the fourth reagent liquid storage tank, and a flow channel is opened on the support part. One end of the flow channel covers the bottom of the drainage hole and is connected to the drainage hole, and the other end of the flow channel is connected to one end of the first channel. The specific structure for realizing the lifting of the magnetic separation module is that a connecting bracket is fixedly connected in the installation shell below the fixed plate, a magnetic separation lifting motor is fixedly connected to the upper part of the connecting bracket, a magnetic separation lifting motor is connected to the magnetic separation lifting motor, a magnetic separation lifting screw is threadedly connected to a lifting nut slidably connected to the connecting bracket, and the electromagnetic separation seat is fixedly connected to the lifting nut.

[0010] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, wherein: a first connecting channel is provided on the control connecting valve body, one end of the first connecting channel can cover the other end of the first channel, and the other end of the first connecting channel covers an end of the second channel away from the inlet and outlet holes.

[0011] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, wherein: the control connecting valve body is also provided with a second connecting channel arranged in the height direction with the first connecting channel, one end of the second connecting channel can cover the second channel, and the reagent box body at the other end of the second connecting channel is provided with a plurality of third connecting channels corresponding to the probe pools one by one, and the end of the third connecting channel away from the second connecting channel can be connected to the corresponding probe pool, and the reagent box body at the end of the probe pool away from the third connecting channel is provided with a liquid outlet channel, and the end of the liquid outlet channel away from the third connecting channel is connected to the amplification chamber, and the liquid outlet channel is arranged on the upper part of the probe pool and the amplification chamber.

[0012] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, the fixed frame is connected with a microfluidic pump that can be raised and lowered, and the suction and pressure tube of the microfluidic pump can be just inserted into the amplification chamber.

[0013] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic card box in the present invention, wherein: the upper end of the fixed frame is slidably connected to a movable beam that can slide left and right, and the movable beam is slidably connected to a vertically arranged first vertical plate that can move forward and backward, and the first vertical plate is slidably connected to a lifting seat, and the stirring magnetic rod is connected to the lifting seat, and the movable beam is fixedly connected to a first movable motor, and the first movable motor is connected to a first movable screw rod, and the first movable screw rod is threadedly connected to the first vertical plate, and the first vertical plate is fixedly connected to a first lifting motor, and the first lifting motor is connected to a first lifting screw rod, and the first lifting screw rod is threadedly connected to the lifting seat.

[0014] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, wherein: the upper end of the fixed frame is fixedly connected to a fixed plate, a first extension plate extending downward is fixedly connected to the fixed plate, a second lifting motor is fixedly connected to the first extension plate, the second lifting motor is connected to a second lifting screw rod, a lifting connecting plate slidably connected to the first extension plate is threadedly connected to the second lifting screw rod, and the microfluidic pump is fixedly connected to the lifting connecting plate.

[0015] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, wherein: the fixed plate is also fixedly connected to a second extension plate extending downward, the second extension plate is slidably connected to a second lifting plate that can be lifted and lowered, the second lifting plate is fixedly connected to a third lifting motor, the third lifting motor is connected to a lifting gear, the second extension plate is fixedly connected to a lifting rack that cooperates with the lifting gear, the second lifting plate is fixedly connected to a rotating motor, the steering motor is connected to a rotating shaft, the lower end of the rotating shaft is fixedly connected to a bidirectional linear drive, the two ends of the bidirectional linear drive are respectively connected to rotating claws that can perform reciprocating linear motion, the outer periphery of the upper part of the control connecting valve body is provided with two plug-in countersunk holes corresponding to the rotating claws one by one, and the rotating claws can be just inserted into the corresponding plug-in countersunk holes.

[0016] As a preferred solution of the fully automatic PCR analyzer based on the microfluidic cartridge in the present invention, wherein: a supporting bracket is fixedly connected in the installation shell below the fixed plate, a lifting bracket that can be lifted and lowered is connected to the supporting bracket, a fixed bracket is fixedly connected in the installation shell, a transmission motor is fixedly connected to the upper side of the fixed bracket, a transmission screw is connected to the transmission motor, the lifting bracket is threadedly connected to the transmission screw, a second moving motor is fixedly connected to the lifting bracket, a second moving screw is connected to the second moving motor, a sliding bracket that can move left and right is slidably connected to the lifting bracket, the second moving screw is threadedly connected to the sliding bracket, a heating module is fixedly connected to the upper side of the sliding bracket, a plurality of heating cavities corresponding to the amplification chambers are opened on the heating module, and the heating module can be attached to the lower side of the corresponding amplification chamber through the heating cavity; a detection probe that can be lifted and lowered, rotated and aligned with the heating module is also connected in the installation shell.

[0017] A method for detecting nucleic acid using a fully automatic PCR analyzer based on a microfluidic cartridge comprises the following steps: The first reagent reservoir, the second reagent reservoir, the third reagent reservoir, the fourth reagent reservoir and the amplification liquid pool are respectively added with a mixture of lysate and magnetic beads, a first cleaning solution, a second cleaning solution, an elution solution and an amplification ball, and different TaqMan probe balls are added to different probe pools; The sample to be tested is injected into the first reagent reservoir, and the stirring magnetic bar is reciprocated in the height direction to fully mix the lysate and the sample, release the nucleic acid, and the magnetic beads specifically capture the nucleic acid and adsorb it on the magnetic bar to complete the lysis; The stirring magnetic bar is lifted up and leaves the first reagent reservoir and moves to above the second reagent reservoir, so that the stirring magnetic bar moves up and down, and the first cleaning is performed under the action of the first cleaning solution to separate the magnetic beads from impurities in the solution; The stirring magnetic bar is lifted up and leaves the second reagent reservoir and moves to the top of the third reagent reservoir, so that the stirring magnetic bar moves up and down, and a second cleaning is performed under the action of the second cleaning solution to further remove impurities remaining on the magnetic beads and ensure the purity of the nucleic acid; The stirring magnetic bar is lifted, so that the stirring magnetic bar moves up and away from the third reagent reservoir and moves to above the fourth reagent reservoir, so that the stirring magnetic bar moves up and down, and the elution is performed under the action of the eluent to dissociate the nucleic acid adsorbed on the magnetic beads, and the magnetic separation module moves up and fits on the lower side of the fourth reagent reservoir. The magnetic separation module is started so that the detached magnetic beads are adsorbed on the inner wall of the fourth reagent reservoir, thereby completing the automated extraction of the nucleic acid; The microfluidic pump is operated with negative pressure to allow the liquid from which the nucleic acid has been extracted in the fourth reagent liquid reservoir to enter the amplification liquid pool through the first channel, the second channel and the inlet and outlet holes in sequence to dissolve the amplification beads; The control connecting valve body is moved downward, and when the first connecting channel covers the second channel, the microfluidic pump is operated with positive pressure, and the microfluidic pump pumps the mixed liquid into the probe pool to dissolve different TaqMan probe beads; When the liquid surface in the probe pool is higher than the height of the lower side of the liquid outlet channel, the liquid after dissolving the corresponding TaqMan probe ball enters the amplification chamber for subsequent amplification reaction.

[0018] The heating module is moved left and right to a position aligned with the amplification chamber, and the heating module is moved up. The heating module is attached to the lower end of the corresponding amplification chamber through the heating cavity. The heating element in the heating module works to heat the amplification chamber to a set temperature. The polymerase gradually adds deoxyribonucleoside triphosphates along the 3' end of the template nucleic acid to synthesize a new nucleic acid chain. When the polymerase synthesizes along the template chain to the probe binding position, it uses its 5'-3' exonuclease activity to cut the probe, so that the fluorescent reporter group is separated from the quenching group, releasing the fluorescent signal, and the heating cavity leaves the amplification chamber to complete a cycle. The detection probe is moved to the same level as the amplification chamber. According to the fluorescent groups of different wavelengths connected to the TapMan probe, the detection probe is rotated according to the fluorescent groups of different wavelengths connected to the TapMan probe, so that the excitation light source of the corresponding wavelength on the detection probe and the detection probe are aligned with the amplification chamber, and the fluorescence intensity of different amplification chambers is detected to complete a cycle; return to the above detection step until the set number of cycles is reached, and the signal detection is completed.

[0019] Compared with the prior art, the present invention has the following technical effects: the present invention can realize the automatic extraction of nucleic acid and facilitate the subsequent amplification through the joint arrangement of several reagent storage tanks, movable stirring magnetic bars and liftable magnetic separation modules; through the joint arrangement of an amplification pool, a control connecting valve body that can be lifted and rotated, a probe pool, an amplification chamber and a liftable microfluidic pump, the sampling, distribution, amplification and signal detection of nucleic acid are further realized, the efficiency of nucleic acid detection is improved, and the situation that the unstable artificial extraction affects the effect of nucleic acid amplification is avoided; the present invention can be applied to the integrated work of nucleic acid extraction, amplification and signal detection. BRIEF DESCRIPTION OF THE DRAWINGS

[0020] In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the following briefly introduces the drawings required for describing the embodiments. Obviously, the drawings described below are only some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without creative labor. Among them: Figure 1 The three-dimensional structure of the present invention Figure 1 .

[0021] Figure 2 This is a three-dimensional structural diagram after the right cover plate and the door body are hidden in the present invention.

[0022] Figure 3 for Figure 2 A partial enlarged view of point A in the middle.

[0023] Figure 4 It is a three-dimensional structural diagram of the reagent reaction box in the present invention.

[0024] Figure 5 It is a top view of the reagent reaction box in the present invention.

[0025] Figure 6 for Figure 5 The view from the center BB.

[0026] Figure 7 for Figure 6 A partial enlarged view of point C in the middle.

[0027] Figure 8 This is a three-dimensional structural diagram of the reagent box body and the control connecting valve body being connected to the reagent reaction box when the reagent box body and the control connecting valve body are set in a transparent state.

[0028] Fig. 9 for Figure 8 A partial enlarged view of point D in the middle.

[0029] Fig.10 for Figure 8 A partial enlarged view of point E in the middle.

[0030] Fig.11 The three-dimensional structure behind the cover plate on the front side of the housing is hidden in the present invention. Figure 1 .

[0031] Fig.12 for Fig.11 A partial enlarged view of point F in the middle.

[0032] Fig.13 The three-dimensional structure behind the cover plate on the front side of the housing is hidden in the present invention. Figure 2 .

[0033] Fig.14 for Fig.13 A partial enlarged view of point G in the middle.

[0034] Fig.15 This is a three-dimensional structural diagram after the cover plate on the rear side of the housing is hidden in the present invention.

[0035] Fig.16 for Fig.15 A partial enlarged view of point H in the middle.

[0036] Fig.17 for Fig.15 A partial enlarged view of point I in the middle.

[0037] In the figure, 100 is a containing component, 101 is a door body, 102 is a right cover plate, 103 is a mounting shell, 104 is a fixed frame, 105 is a mounting plate, 200 is a nucleic acid extraction component, 201 is an electromagnetic separation seat, 201a is a magnetic separation port, 202 is a first lifting screw rod, 203 is a stirring sleeve, 204 is a stirring magnetic rod, 205 is a lifting seat, 206 is a first lifting motor, 207 is a first vertical plate, 208 is a first moving screw rod, 209 is a moving beam, 210 is a first moving motor, 211 is a lifting block, 212 is a third lifting screw rod, 213 is a control connecting valve body, 213a is a first connecting channel, 213b is a second connecting channel, 213c is a plug-in countersunk hole, 214 is a fixed plate, 215 is a magnetic separation lifting screw rod, 216 is a magnetic separation lifting motor, 217 is a connecting bracket, 218 is a lifting nut, 300 is a reagent reaction box, 301 is a reagent box body, 301a is an amplification liquid pool, 301b first reagent reservoir, 301c second reagent reservoir, 301d third reagent reservoir, 301e fourth reagent reservoir, 301e-1 drainage hole, 301g second channel, 301h third connecting channel, 301i probe pool, 301j amplification chamber, 301k liquid outlet channel, 302 bottom plate, 302a support part, 302a-1 flow channel, 302a-2 first channel, 400 nucleic acid detection component, 401 lifting rack, 402 lifting gear, 403 second lifting plate, 404 third lifting motor, 405 lifting connecting plate, 406 microfluidic pump, 407 07 steering motor, 408 rotating shaft, 409 bidirectional linear drive, 410 second extension plate, 411 sliding bracket, 412 lower detection screw, 413 lower detection motor, 414 detection angle adjustment motor, 415 upper detection screw, 416 upper detection motor, 417 detection plate, 418 detection probe, 419 heating module, 419a heating chamber, 420 transmission screw, 421 second moving screw, 422 lifting bracket, 423 transmission motor, 424 fixed bracket, 425 second moving motor, 426 first extension plate, 427 second lifting motor, 428 second lifting screw. DETAILED DESCRIPTION

[0038] In order to make the above-mentioned objects, features and advantages of the present invention more obvious and easy to understand, the specific implementation methods of the present invention are described in detail below in conjunction with the accompanying drawings.

[0039] In the following description, many specific details are set forth to facilitate a full understanding of the present invention, but the present invention may also be implemented in other ways different from those described herein, and those skilled in the art may make similar generalizations without violating the connotation of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0040] Secondly, the term "one embodiment" or "embodiment" as used herein refers to a specific feature, structure, or characteristic that may be included in at least one implementation of the present invention. The term "in one embodiment" that appears in different places in this specification does not necessarily refer to the same embodiment, nor does it refer to a separate or selective embodiment that is mutually exclusive with other embodiments.

[0041] Example 1 Reference Figures 1 to 9 , this embodiment provides a fully automatic PCR analyzer based on a microfluidic cartridge, which can realize automated extraction of nucleic acids and facilitate amplification.

[0042] A fully automatic PCR analyzer based on a microfluidic cartridge includes a containing component 100, the containing component 100 includes a mounting shell 103 with a containing cavity, a right cover plate 102 is fixedly connected to the right side of the mounting shell 103, an openable and closable door body 101 is connected to the right cover plate 102, a fixed frame 104 is fixedly connected inside the mounting shell 103, a mounting plate 105 is fixed to the fixed frame 104, and a plug-in sink groove is provided on the side of the mounting plate 105 opposite to the door body 101.

[0043] Specifically, a reagent reaction box 300 is connected to the installation shell 103, and the lower end of the reagent reaction box 300 can be inserted into the installation shell 103 through the plug-in sink. The reagent kit includes a bottom plate 302, and a reagent kit body 301 is fixedly connected to the upper side of the bottom plate 302. The reagent kit body 301 is provided with a channel communication control hole, and the upper side of the reagent kit body 301 outside one end of the channel communication control hole in the left and right directions has a plurality of reagent storage tanks and amplification liquid tanks 301a; a nucleic acid extraction component 200 is provided above the installation plate 105, and the nucleic acid extraction component 200 includes a stirring magnetic bar 204 that can be lifted and moved in the front and rear directions and the left and right directions, and the stirring magnetic bar 204 can extend into any reagent storage tank, and a magnetic separation module that can be lifted and lowered is connected to the fixed frame 104 below the installation plate 105, and the magnetic separation module includes an electromagnetic separation seat 201, and the upper end of the electromagnetic separation seat 201 is provided with a magnetic separation port 201a corresponding to the fourth reagent storage tank 301e, and the electromagnetic separation port 201a at the bottom of the magnetic separation port 201a is provided with a magnetic separation port 201a. The separation seat 201 is connected to an electromagnet (which is a prior art). A support portion 302a for facilitating the flow of liquid is fixed on the bottom plate 302 at the bottom of the fourth reagent storage tank 301e. A flow channel 302a-1 is provided on the support portion 302a. One end of the flow channel 302a-1 covers the drainage hole 301e-1 provided at the bottom of the fourth reagent storage tank 301e. One end of the flow channel 302a-1 is connected to the drainage hole 301e-1, and the other end of the flow channel 302a-1 is connected to the fourth reagent storage tank 301e. One end of a channel 302a-2 is connected; the specific structure for realizing the lifting of the magnetic separation module is that the lower part of the installation shell 103 is fixedly connected to a connecting bracket 217, the upper part of the connecting bracket 217 is fixedly connected to a magnetic separation lifting motor 216, the magnetic separation lifting motor 216 is connected to a magnetic separation lifting screw rod 215, the magnetic separation lifting screw rod 215 is threadedly connected to a lifting nut 218 slidably connected to the connecting bracket 217, and the electromagnetic separation seat 201 is fixedly connected to the lifting nut 218.

[0044] The first reagent reservoir 301b, the second reagent reservoir 301c, the third reagent reservoir 301d and the fourth reagent reservoir 301e are respectively added with a mixture of lysate and magnetic beads, a first cleaning solution, a second cleaning solution and an elution solution; the sample to be tested is injected into the first reagent reservoir 301b, and the stirring magnetic bar 204 is reciprocated in the height direction to fully mix the lysate with the sample and release the nucleic acid. The magnetic beads specifically capture the nucleic acid and adsorb on the magnetic bar to complete the lysis; the stirring magnetic bar 204 is lifted up and leaves the first reagent reservoir 301b and moves to the top of the second reagent reservoir 301c, so that the stirring magnetic bar 204 moves up and down, and the first cleaning is performed under the action of the first cleaning solution, and the magnetic beads are separated from the impurities in the solution; the stirring magnetic bar 204 is lifted up and leaves the first reagent reservoir 301b and moves to the top of the second reagent reservoir 301c, and the stirring magnetic bar 204 is moved up and down, and the first cleaning is performed under the action of the first cleaning solution, and the magnetic beads are separated from the impurities in the solution; the stirring magnetic bar 204 is 04 is lifted up and leaves the second reagent reservoir 301c and moves to above the third reagent reservoir 301d, so that the stirring magnetic bar 204 moves up and down, and a second cleaning is performed under the action of the second cleaning solution to further remove impurities remaining on the magnetic beads and ensure the purity of the nucleic acid; the stirring magnetic bar 204 is lifted up and moves up and leaves the third reagent reservoir 301d and moves to above the fourth reagent reservoir 301e, so that the stirring magnetic bar 204 moves up and down, and elution is performed under the action of the elution solution to dissociate the nucleic acid adsorbed on the magnetic beads, the magnetic separation module moves up and fits on the lower side of the fourth reagent reservoir 301e, and the magnetic separation module is started so that the detached magnetic beads are adsorbed on the inner wall of the fourth reagent reservoir 301e, thereby completing the automated extraction of nucleic acid.

[0045] The process of realizing the lifting of the magnetic separation module is to control the movement of the magnetic separation lifting motor 216, rotate the magnetic separation lifting screw 215, drive the lifting nut 218 to move in the height direction, and drive the lifting nut 218 to move the electromagnetic separation seat 201. The movement direction of the magnetic separation lifting motor 216 is controlled to move the electromagnetic separation seat 201 to the required height direction. When the electromagnetic separation seat 201 is lifted to the set height, the magnetic separation lifting motor 216 stops moving.

[0046] Specifically, the upper end of the fixed frame 104 is slidably connected to a moving beam 209 that can slide left and right, and the moving beam 209 is slidably connected to a vertically arranged first vertical plate 207 that can move forward and backward, and the first vertical plate 207 is slidably connected to a lifting seat 205, and the stirring magnetic rod 204 is connected to the lifting seat 205. The moving beam 209 is fixedly connected to a first moving motor 210, and the first moving motor 210 is connected to a first moving screw rod 208, and the first moving screw rod 208 is threadedly connected to the first vertical plate 207, and the first vertical plate 207 is fixedly connected to a first lifting motor 206, and the first lifting motor 206 is connected to a first lifting screw rod 202, and the first lifting screw rod 202 is threadedly connected to the lifting seat 205; the first vertical plate 207 is also slidably connected to There is a lifting block 211, and a fourth lifting motor is fixedly connected to the upper end of the first vertical plate 207. The fourth lifting motor is connected to a third lifting screw rod 212 that is rotatably connected between the first vertical plate 207 and the mounting plate 105. The third lifting screw rod 212 is threadedly connected to the lifting block 211. The lifting block 211 is clamped with a stirring sleeve 203 through a snap ring (the snap ring is a prior art, as long as it can clamp the round object and prevent it from falling. It is not an improvement point of the present invention, and the specific structure of the snap ring is not repeated). The electromagnetic stirring rod can extend downward into the stirring sleeve 203. The stirring sleeve 203 can prevent the electromagnetic stirring rod from directly contacting the reagent. The stirring sleeve 203 is a disposable item. After each test, the stirring sleeve 203 is removed from the snap ring and a new stirring sleeve 203 is installed.

[0047] A third moving motor is also fixedly connected above the fixed frame 104, and a moving nut that can move left and right is also slidably connected above the fixed frame 104. The end of the moving nut away from the third moving motor is fixedly connected to the moving beam 209, and the third moving motor is connected to a third moving screw rod, and the third moving screw rod and the moving nut are threadedly connected.

[0048] The process of realizing the movement of the stirring magnetic rod 204 in the left and right, front and back, and height directions is as follows: when the stirring magnetic rod 204 needs to be moved in the front and back directions, the first moving motor 210 is activated, the first moving screw rod 208 rotates, and drives the first vertical plate 207 to slide along the front and back directions of the moving beam 209; when the stirring magnetic rod 204 moves to a suitable position in the front and back directions, the first moving motor 210 stops; when the position of the stirring magnetic rod 204 in the height direction needs to be adjusted, the first lifting motor 206 and the fourth lifting motor are activated at the same time, the first lifting screw rod 202 and the third lifting screw rod 212 rotate, the first lifting screw rod 202 and the third lifting screw rod 212 respectively drive the lifting seat 205 and the lifting block 211 to move in the height direction, and the lifting The seat 205 and the lifting block 211 respectively drive the stirring magnetic rod 204 and the stirring sleeve 203 to rise and fall. When the stirring sleeve 203 extends into the corresponding reagent storage tank, the stirring magnetic rod 204 extends into a section of the stirring magnetic rod 204 in the stirring sleeve 203 and contacts the reagent. When the stirring magnetic rod 204 needs to move up and down reciprocatingly, the movement direction of the first lifting motor 206 and the fourth lifting motor is controlled to make the stirring sleeve 203 and the stirring magnetic rod 204 move up and down reciprocatingly; when the stirring magnetic rod 204 is lifted to a suitable position and the position in the height direction does not need to be adjusted, the first lifting motor 206 and the fourth lifting motor stop moving; when the position of the stirring magnetic rod 204 in the left and right directions needs to be adjusted to achieve the switching of the third reagent storage tank 301d to the direction where the fourth reagent storage tank 301e is located, the third moving motor is controlled to move, the third moving screw rod rotates, the third moving screw rod drives the moving nut to slide along the left and right directions of the fixed frame 104, and the moving nut drives the moving beam 209 to move in the left and right directions. When the stirring magnetic rod 204 moves left and right to a suitable position, the third moving motor stops moving.

[0049] Specifically, it also includes a nucleic acid detection component 400, a fixed plate 214 is fixedly connected to the upper end of the fixed frame 104, a third movable motor is fixedly connected to the fixed plate 214, the nucleic acid detection component 400 includes a first extension plate 426 fixedly connected to the fixed plate 214 and extending downward, a second lifting motor 427 is fixedly connected to the first extension plate 426, the second lifting motor 427 is connected to a second lifting screw rod 428, the second lifting screw rod 428 is threadedly connected to a lifting connecting plate 405 slidably connected to the first extension plate 426, and a microfluidic pump 406 is fixedly connected to the lifting connecting plate 405.

[0050] The action process of realizing the lifting of the microfluidic pump 406 is to control the second lifting motor 427 to move, the second lifting screw 428 to rotate, and the second lifting screw 428 drives the lifting connecting plate 405 to move in the height direction. When the microfluidic pump 406 is driven to a suitable height, the second lifting motor 427 stops moving.

[0051] Specifically, the fixed plate 214 is also fixedly connected to a second extension plate 410 extending downward, and a second lifting plate 403 that can be raised and lowered is slidably connected to the second extension plate 410, and a third lifting motor 404 is fixedly connected to the second lifting plate 403, and a lifting gear 402 is connected to the third lifting motor 404, and a lifting rack 401 that cooperates with the lifting gear 402 is fixedly connected to the second extension plate 410, and a rotating motor is fixedly connected to the second lifting plate 403, and a rotating shaft 408 is connected to the steering motor 407, and a bidirectional linear driver 409 is fixedly connected to the lower end of the rotating shaft 408, and rotating claws that can perform reciprocating linear motion are respectively connected to the two ends of the bidirectional linear driver 409, and two plug-in countersunk holes 213c corresponding to the rotating claws are opened on the outer periphery of the upper part of the control connecting valve body 213, and the rotating claws can just be inserted into the corresponding plug-in countersunk holes 213c.

[0052] In the initial state, one side of the two rotating claws that are relatively arranged is plugged into the corresponding plug-in countersunk hole 213c; before the detection is completed, it is always plugged into the corresponding plug-in countersunk hole 213c; the action process of controlling the rotation of the connecting valve body 213 and adjusting the position in the height direction is that when the connecting valve body 213 is controlled to be lifted to a suitable height and the channel connectivity needs to be adjusted, the rotating motor is activated, the rotating shaft 408 rotates, the rotating shaft 408 drives the bidirectional linear driver 409 to rotate, and the bidirectional linear driver 409 drives the connecting valve body 213 to rotate through the two rotating claws. When the connecting valve body 213 is rotated to the required angle, the rotating motor stops; when the height of the connecting valve body 213 needs to be adjusted, the third lifting motor 404 is activated, the lifting gear 402 rotates, the lifting gear 402 rolls along the lifting rack 401, and the lifting gear 402 drives the second lifting plate 403 to lift and lower. When the connecting valve body 213 is controlled to be lifted and lowered to a suitable height, the third lifting motor 404 stops.

[0053] This embodiment can realize the automatic extraction of nucleic acid and facilitate the subsequent amplification by jointly setting a plurality of reagent storage tanks, a movable stirring magnetic rod 204 and a lifting magnetic separation module.

[0054] Example 2 Reference Figure 8 to Figure 10 This embodiment provides a fully automatic PCR analyzer based on a microfluidic cartridge. The difference between this embodiment and embodiment 1 is that it can further realize the injection and distribution of nucleic acids.

[0055] Specifically, the upper part of the reagent box body 301 at the other end of the channel communication control hole in the left-right direction has a plurality of probe pools 301i that can be connected to the amplification liquid pool 301a, and the probe pool 301i has an amplification chamber 301j on the reagent box body 301 away from the channel communication hole at one end in the left-right direction. A microfluidic pump 406 that can be raised and lowered is connected to the fixed frame 104, and the suction and pressure pipe of the microfluidic pump 406 can be just inserted into the amplification chamber 301j. The reagent box body 301 is just movably connected to the control communication valve body 213 through the channel communication control hole. The plurality of reagent storage pools are respectively a first reagent box, a second reagent box, and a third reagent box. The fourth reagent reservoir 301e has a drainage hole 301e-1 at the bottom, and a first channel 302a-2 is formed on the bottom plate 302 below the drainage hole 301e-1. The bottom of the amplification liquid reservoir 301a has an inlet and outlet hole, and a second channel 301g is formed on the reagent kit body 301 below the inlet and outlet hole. The other end of the first channel 302a-2 (the other end of the first channel 302a-2 is the end away from the drainage hole 301e-1) The control connecting valve body 213 is provided with a first connecting channel 213a, one end of the first connecting channel 213a can cover the other end of the first channel 302a-2, and the other end of the first connecting channel 213a covers the end of the second channel 301g away from the inlet and outlet holes. The control connecting valve body 213 is also provided with a second connecting channel 213b spaced apart from the first connecting channel 213a in the height direction, one end of the second connecting channel 213b can cover the second channel 301g, and the second connecting channel 213b can cover the second channel 301g. A plurality of third connecting channels 301h corresponding to the probe pools 301i are provided on the reagent box body 301 at the other end of the channel 213b. The end of the third connecting channel 301h away from the second connecting channel 213b can be connected to the corresponding probe pool 301i. A liquid outlet channel 301k is provided on the reagent box body 301 at the end of the probe pool 301i away from the third connecting channel 301h. The end of the liquid outlet channel 301k away from the third connecting channel 301h is connected to the amplification chamber 301j. The liquid outlet channel 301k is arranged at the upper part of the probe pool 301i and the amplification chamber 301j.

[0056] The control connecting valve body 213 is rotated in the amplification liquid pool 301a. When one end of the first connecting channel 213a just covers the other end of the first channel 302a-2, and the other end of the first connecting channel 213a covers the end of the second channel 301g away from the inlet and outlet holes, the control connecting valve body 213 stops rotating, the microfluidic pump 406 moves downward, the suction and pressure tube is inserted into the amplification chamber 301j, the microfluidic pump 406 is activated, and the action direction of the microfluidic pump 406 is controlled to suck the liquid in the fourth reagent reservoir 301e, so that the purified nucleic acid in the fourth reagent reservoir 301e is sucked into the amplification liquid pool 301a to dissolve the amplification ball; the control connecting valve body 213 is moved downward, and when the second connecting channel 213b When the height moves down to the point where the second connecting channel 213b and the second channel 301g are at the same height, the control connecting valve body 213 stops rising and falling, and the control connecting valve body 213 rotates. When one end of the second connecting channel 213b just covers the second channel 301g, the microfluidic pump 406 reverses. The positive pressure operation of the microfluidic pump 406 sequentially passes the mixed liquid through the second channel 301g, the second connecting channel 213b and several third connecting channels 301h into different probe pools 301i to dissolve different TaqMan probe balls. The liquid that has dissolved the TaqMan probe balls is continued to be pressed into the amplification chamber 301j through the liquid outlet channel 301k to further achieve amplification.

[0057] Example 3 Reference Figures 11 to 17 This embodiment provides a fully automatic PCR analyzer based on a microfluidic cartridge. The difference between this embodiment and embodiments 1 and 2 is that it can further realize signal detection.

[0058] Specifically, a support bracket is fixedly connected in the installation shell 103 below the fixed plate 214, and a lifting bracket 422 capable of lifting and lowering is connected to the support bracket. A fixed bracket 424 is fixedly connected in the installation shell 103, and a transmission motor 423 is fixedly connected to the upper side of the fixed bracket 424, and a transmission screw 420 is connected to the transmission motor 423. The lifting bracket 422 is threadedly connected to the transmission screw 420, and a second moving motor 425 is fixedly connected to the lifting bracket 422. The second moving motor 425 A second movable screw rod 421 is connected to the upper part, and a sliding bracket 411 that can move left and right is slidably connected to the lifting bracket 422. The second movable screw rod 421 and the sliding bracket 411 are threadedly connected. A heating module 419 is fixedly connected to the upper side of the sliding bracket 411. The heating module 419 has a plurality of heating cavities 419a corresponding to the amplification chambers 301j. The heating module 419 can be attached to the lower side of the corresponding amplification chamber 301j through the heating cavity 419a. The heating cavity 419a of the heating module 419 An electronic heating element is provided inside (preferably a metal heating block, and the metals commonly used in PCR are generally copper, aluminum and other metals with good thermal conductivity, which are prior art); a detection probe 418 that can be lifted, rotated and aligned with the heating module 419 is also connected to the mounting shell 103, and the structure for realizing the movement of the detection probe 418 is specifically as follows: the left part of the sliding bracket 411 is slidably connected to a connecting frame that can move forward and backward, the left end of the sliding bracket 411 is fixedly connected to a lower detection motor 413, the lower detection motor 413 is connected to a lower detection screw rod 412, the lower detection screw rod 412 is threadedly connected to a connecting frame that is slidably connected to the sliding bracket 411 and can move forward and backward, the upper end of the connecting frame is fixedly connected to an upper detection motor 416, the upper detection motor 416 is connected to an upper detection screw rod 415, the upper detection screw rod 415 is threadedly connected to a detection plate 417 that is slidably connected to the connection frame, the left end of the detection plate 417 is fixedly connected to a detection angle adjustment motor 414, and the detection angle adjustment motor 414 is connected to a detection probe 418.

[0059] The position adjustment process of the detection probe 418 in the left and right, front and back, and angle is as follows: when the position of the detection probe 418 and the heating module 419 in the left and right directions needs to be adjusted, the second moving motor 425 is actuated, the second moving screw 421 rotates, the second moving screw 421 drives the sliding bracket 411 to move, and the sliding bracket 411 drives the heating module 419 and the detection probe 418 to move in the left and right directions. When the heating module 419 moves to a suitable position left and right, the second moving motor 425 stops; when the height position of the heating module 419 needs to be adjusted, the transmission motor 423 is actuated, the transmission screw 420 rotates, and the transmission screw 420 drives the lifting bracket 422 to move up and down. The lifting bracket 422 drives the heating module 419 to rise and fall via the sliding bracket 411. When the heating module 419 is lifted to a suitable position, the transmission motor 423 stops working; when it is necessary to adjust the height of the detection probe 418, the upper detection motor 416 is activated, the upper detection screw rod 415 rotates, and the upper detection screw rod 415 drives the detection probe 418 to rise and fall via the detection plate 417. When the detection probe 418 is lifted to a suitable position, the upper detection motor 416 stops working; when it is necessary to adjust the angle of the detection probe 418, the detection angle adjustment motor 414 is activated to drive the detection probe 418 to rotate. When the detection probe 418 rotates to a suitable angle, the detection angle adjustment motor 414 stops working.

[0060] During detection, the heating module 419 is moved left and right to the position aligned with the amplification chamber 301j, and the heating module 419 is moved up. The heating module 419 is attached to the lower end of the corresponding amplification chamber 301j through the heating chamber 419a. The heating element in the heating module 419 works to heat the amplification chamber 301j to the set temperature. The polymerase gradually adds deoxyribonucleoside triphosphates along the 3' end of the template nucleic acid to synthesize a new nucleic acid chain. When the polymerase synthesizes along the template chain to the probe binding position, it uses its 5'-3' exonuclease activity to cut the probe, so that the fluorescent reporter group is separated from the quenching group, and the fluorescent signal is released, so that the heating The cavity 419a leaves the amplification chamber 301j, completing one cycle. The detection probe 418 moves to the same level as the amplification chamber 301j, and according to the fluorescent groups of different wavelengths connected to the TaqMan probe, the detection probe 418 is rotated so that the excitation light source of the corresponding wavelength on the detection probe 418 and the detection probe 418 are aligned with the amplification chamber 301j, and the fluorescence intensity of different amplification chambers 301j is detected to complete one cycle; the above detection step is returned until the set number of cycles is reached to complete the signal detection.

[0061] After the detection is completed, the rotating motor is activated to make the first connecting channel 213a offset from the second channel 301g, the rotating motor stops, the bidirectional linear drive 409 is activated to make the rotating claw leave the detection connecting valve body, the third lifting motor 404 is activated to move the rotating claw up to the set height, and the third lifting motor 404 stops; open the door body 101, remove the stirring sleeve 203, loosen the screws connecting the reagent box body 301 and the fixed plate 214, remove the reagent box body 301, insert a new stirring sleeve 203, insert the new reagent box body 301 into the fixed plate 214 through the plug-in sink (a new control connecting valve body 213 is plugged in the channel connecting control hole), use screws to fix the reagent box body 301 to the fixed plate 214, and wait for the next nucleic acid test.

[0062] Example 4 Reference Figure 3 and Figure 6 This embodiment provides a fully automatic PCR analyzer based on a microfluidic cartridge. The difference between this embodiment and embodiment 3 is that it can further realize nucleic acid signal detection.

[0063] Specifically, the method for detecting nucleic acid using a fully automatic PCR analyzer based on a microfluidic cartridge comprises the following steps: The first reagent reservoir 301b, the second reagent reservoir 301c, the third reagent reservoir 301d, the fourth reagent reservoir 301e and the amplification liquid pool 301a are respectively added with a mixture of lysate and magnetic beads, a first cleaning solution, a second cleaning solution, an eluent and an amplification ball, and different TaqMan probe balls are added to different probe pools 301i; The sample to be tested is injected into the first reagent reservoir 301b, and the stirring magnetic bar 204 is reciprocated in the height direction to fully mix the lysate with the sample, release the nucleic acid, and the magnetic beads specifically capture the nucleic acid and adsorb it on the magnetic bar to complete the lysis; The stirring magnetic bar 204 is lifted up and leaves the first reagent reservoir 301b and moves to above the second reagent reservoir 301c, so that the stirring magnetic bar 204 moves up and down, and the first cleaning is performed under the action of the first cleaning solution to separate the magnetic beads from the impurities in the solution; The stirring magnetic bar 204 is lifted up and leaves the second reagent reservoir 301c and moves to above the third reagent reservoir 301d, so that the stirring magnetic bar 204 moves up and down, and a second cleaning is performed under the action of the second cleaning solution to further remove impurities remaining on the magnetic beads and ensure the purity of the nucleic acid; The stirring magnetic bar 204 is lifted, so that the stirring magnetic bar 204 moves upward away from the third reagent storage tank 301d and moves to above the fourth reagent storage tank 301e, so that the stirring magnetic bar 204 moves up and down, and the elution is performed under the action of the eluent to dissociate the nucleic acid adsorbed on the magnetic beads, and the magnetic separation module moves upward to fit the lower side of the fourth reagent storage tank 301e. The magnetic separation module is started so that the detached magnetic beads are adsorbed on the inner wall of the fourth reagent storage tank 301e, and the automatic extraction of nucleic acid is completed; The microfluidic pump 406 is operated with negative pressure to move the liquid from which the nucleic acid has been extracted in the fourth reagent reservoir 301e into the amplification liquid reservoir 301a through the first channel 302a-2, the second channel 301g and the inlet and outlet holes in sequence to dissolve the amplification beads; The control connecting valve body 213 is moved downward, and when the first connecting channel 213a covers the second channel 301g, the microfluidic pump 406 is operated with positive pressure, and the microfluidic pump 406 pumps the mixed liquid into the probe pool 301i to dissolve different TaqMan probe beads; When the liquid surface in the probe pool 301i is higher than the height of the lower side of the liquid outlet channel 301k, the liquid after dissolving the corresponding TaqMan probe ball enters the amplification chamber 301j for subsequent amplification reaction.

[0064] It should be noted that the above embodiments are only used to illustrate the technical solutions of the present invention rather than to limit it. Although the present invention has been described in detail with reference to the preferred embodiments, those skilled in the art should understand that the technical solutions of the present invention may be modified or replaced by equivalents without departing from the spirit and scope of the technical solutions of the present invention, which should all be included in the scope of the claims of the present invention.

Claims

1. A fully automatic PCR analyzer based on a microfluidic cartridge, characterized in that: These include, A mounting shell (103) having a receiving cavity, wherein one side of the mounting shell (103) in the left-right direction is connected to a door body (101) that can be opened and closed, a fixed frame (104) is fixedly connected inside the mounting shell (103), a mounting plate (105) is fixed on the fixed frame (104), and a plug-in recess is formed on one side of the mounting plate (105) that is opposite to the door body (101); A reagent reaction box (300), wherein the lower end of the reagent reaction box (300) can be inserted into the mounting housing (103) through a plug-in sink, the reagent box comprises a bottom plate (302), a reagent box body (301) is fixedly connected to the upper side of the bottom plate (302), a channel communication control hole is provided on the reagent box body (301), a plurality of reagent liquid storage pools and amplification liquid pools (301a) are provided on the upper side of the reagent box body (301) outside one end of the channel communication control hole in the left-right direction, a plurality of probe pools (301i) that can be connected to the amplification liquid pools (301a) are provided on the upper part of the reagent box body (301) at the other end of the channel communication control hole in the left-right direction, and the probe pools (301i) have an amplification chamber (301j) on the reagent box body (301) away from one end of the channel communication hole in the left-right direction; A nucleic acid extraction component (200) is arranged in a mounting shell (103), and the nucleic acid extraction component (200) includes a stirring magnetic bar (204) that can be raised and lowered and can be moved in the front-to-back direction and the left-to-right direction. The stirring magnetic bar (204) can extend into any reagent storage tank. A magnetic separation module that can be raised and lowered is connected to the fixed frame (104) below the mounting plate (105).

2. The fully automatic PCR analyzer based on the microfluidic cartridge according to claim 1, characterized in that: The reagent box body (301) is movably connected to a control communication valve body (213) via a channel communication control hole. The plurality of reagent storage tanks are respectively a first reagent storage tank (301b), a second reagent storage tank (301c), a third reagent storage tank (301d) and a fourth reagent storage tank (301e). The fourth reagent storage tank (301e) has a drainage hole (301e-1) at the bottom. The reagent box body (301) below the drainage hole (301e-1) has a first channel (302a-2). The bottom of the amplification liquid pool (301a) has a liquid inlet and outlet hole. The reagent box body (301) below the liquid inlet and outlet hole has a second channel (301g). The other end of the first channel (302a-2) can be connected to the amplification liquid pool (301a) via the second channel (301g) and the liquid inlet and outlet hole in sequence.

3. The fully automatic PCR analyzer based on the microfluidic cartridge according to claim 2, characterized in that: The control connecting valve body (213) is provided with a first connecting channel (213a), one end of the first connecting channel (213a) can cover the other end of the first channel (302a-2), and the other end of the first connecting channel (213a) covers an end of the second channel (301g) away from the liquid inlet and outlet holes.

4. The fully automatic PCR analyzer based on the microfluidic cartridge according to claim 2, characterized in that: The control connecting valve body (213) is also provided with a second connecting channel (213b) spaced apart from the first connecting channel (213a) in the height direction, one end of the second connecting channel (213b) can cover the second channel (301g), and the reagent box body (301) at the other end of the second connecting channel (213b) is provided with a plurality of third connecting channels (301h) corresponding to the probe pools (301i) one by one, and the end of the third connecting channel (301h) away from the second connecting channel (213b) can be connected to the corresponding probe pool (301i), and the reagent box body (301) at the end of the probe pool (301i) away from the third connecting channel (301h) is provided with a liquid outlet channel (301k), and the end of the liquid outlet channel (301k away from the third connecting channel (301h) is connected to the amplification chamber (301j), and the liquid outlet channel (301k) is arranged at the upper part of the probe pool (301i) and the amplification chamber (301j).

5. The fully automatic PCR analyzer based on the microfluidic cartridge according to any one of claims 2 to 4, characterized in that: The fixed frame (104) is connected to a microfluidic pump (406) that can be raised and lowered, and the suction and pressure tube of the microfluidic pump (406) can be just inserted into the amplification chamber (301j).

6. The fully automatic PCR analyzer based on the microfluidic cartridge according to any one of claims 1 to 4, characterized in that: The upper end of the fixed frame (104) is slidably connected to a movable beam (209) capable of sliding left and right, the movable beam (209) is slidably connected to a vertically arranged first vertical plate (207) capable of moving forward and backward, the first vertical plate (207) is slidably connected to a lifting seat (205), the stirring magnetic rod (204) is connected to the lifting seat (205), the movable beam (209) is fixedly connected to a first movable motor (210), the first movable motor (210) is connected to a first movable screw rod (208), the first movable screw rod (208) and the first vertical plate (207) are threadedly connected, the first vertical plate (207) is fixedly connected to a first lifting motor (206), the first lifting motor (206) is connected to a first lifting screw rod (202), and the first lifting screw rod (202) and the lifting seat (205) are threadedly connected.

7. The fully automatic PCR analyzer based on the microfluidic cartridge according to claim 5, characterized in that: The upper end of the fixed frame (104) is fixedly connected to a fixed plate (214), a first extension plate (426) extending downward is fixedly connected to the fixed plate (214), a second lifting motor (427) is fixedly connected to the first extension plate (426), the second lifting motor (427) is connected to a second lifting screw rod (428), the second lifting screw rod (428) is threadedly connected to a lifting connecting plate (405) slidably connected to the first extension plate (426), and the microfluidic pump (406) is fixedly connected to the lifting connecting plate (405).

8. The fully automatic PCR analyzer based on the microfluidic cartridge according to claim 5, characterized in that: The fixed plate (214) is also fixedly connected to a second extension plate (410) extending downward, the second extension plate (410) is slidably connected to a second lifting plate (403) capable of being raised and lowered, the second lifting plate (403) is fixedly connected to a third lifting motor (404), the third lifting motor (404) is connected to a lifting gear (402), the second extension plate (410) is fixedly connected to a lifting rack (401) matched with the lifting gear (402), and the second lifting plate (403) is fixedly connected to a rotating motor, and a rotating shaft (408) is connected to the steering motor (407). The lower end of the rotating shaft (408) is fixedly connected to a bidirectional linear driver (409). The two ends of the bidirectional linear driver (409) are respectively connected to rotating claws capable of reciprocating linear motion. The outer periphery of the upper part of the control connecting valve body (213) is provided with two plug-in countersunk holes (213c) corresponding to the rotating claws one by one, and the rotating claws can be just inserted into the corresponding plug-in countersunk holes (213c).

9. The fully automatic PCR analyzer based on the microfluidic cartridge according to claim 8, characterized in that: A support bracket is fixedly connected in the installation shell (103) below the fixed plate (214), and a lifting bracket (422) capable of lifting and lowering is connected to the support bracket. A fixed bracket (424) is fixedly connected in the installation shell (103), and a transmission motor (423) is fixedly connected to the upper side of the fixed bracket (424), and a transmission screw (420) is connected to the transmission motor (423). The lifting bracket (422) is threadedly connected to the transmission screw (420), and a second moving motor (425) is fixedly connected to the lifting bracket (422), and the second moving motor (425) is connected to the second moving screw (420). 1), a sliding bracket (411) capable of moving left and right is slidably connected to the lifting bracket (422), the second movable screw rod (421) and the sliding bracket (411) are threadedly connected, a heating module (419) is fixedly connected to the upper side of the sliding bracket (411), and the heating module (419) is provided with a plurality of heating cavities (419a) corresponding to the amplification chambers (301j) one by one, and the heating module (419) can be attached to the lower side of the corresponding amplification chamber (301j) through the heating cavities (419a); a detection probe (418) capable of lifting, rotating and aligning with the heating module (419) is also connected to the installation shell (103).

10. A method for detecting nucleic acid using the fully automatic PCR analyzer based on the microfluidic cartridge according to claim 9, characterized in that: The following steps are included: The first reagent reservoir (301b), the second reagent reservoir (301c), the third reagent reservoir (301d), the fourth reagent reservoir (301e) and the amplification liquid pool (301a) are respectively added with a mixture of lysate and magnetic beads, a first cleaning solution, a second cleaning solution, an eluent and amplification beads, and different TaqMan probe beads are added to different probe pools (301i); The sample to be tested is injected into the first reagent storage tank (301b), and the stirring magnetic rod (204) is made to reciprocate in the height direction to fully mix the lysate with the sample, release the nucleic acid, and the magnetic beads specifically capture the nucleic acid and adsorb it on the magnetic rod to complete the lysis; The stirring magnetic bar (204) is lifted up and away from the first reagent storage tank (301b) and moved to above the second reagent storage tank (301c), so that the stirring magnetic bar (204) moves up and down, and a first cleaning is performed under the action of the first cleaning solution, so that the magnetic beads are separated from impurities in the solution; The stirring magnetic bar (204) is lifted up and away from the second reagent storage tank (301c) and moved to above the third reagent storage tank (301d), so that the stirring magnetic bar (204) moves up and down, and a second cleaning is performed under the action of the second cleaning solution to further remove impurities remaining on the magnetic beads and ensure the purity of the nucleic acid; The stirring magnetic bar (204) is lifted, so that the stirring magnetic bar (204) moves upward and away from the third reagent storage tank (301d) and moves to above the fourth reagent storage tank (301e), and the stirring magnetic bar (204) moves up and down, and elution is performed under the action of the eluent to dissociate the nucleic acid adsorbed on the magnetic beads, and the magnetic separation module moves upward and fits on the lower side of the fourth reagent storage tank (301e). The magnetic separation module is started so that the detached magnetic beads are adsorbed on the inner wall of the fourth reagent storage tank (301e), thereby completing the automated extraction of the nucleic acid; The microfluidic pump (406) is operated with negative pressure to allow the liquid from which the nucleic acid has been extracted in the fourth reagent liquid reservoir (301e) to enter the amplification liquid reservoir (301a) through the first channel (302a-2), the second channel (301g) and the inlet and outlet holes in sequence to dissolve the amplification beads; The control connecting valve body (213) is moved downward, and when the first connecting channel (213a) covers the second channel (301g), the microfluidic pump (406) is operated with positive pressure, and the microfluidic pump (406) pumps the mixed liquid into the probe pool (301i) to dissolve different TaqMan probe beads; When the liquid surface in the probe pool (301i) is higher than the height of the lower side of the liquid outlet channel (301k), the liquid after dissolving the corresponding TaqMan probe ball enters the amplification chamber (301j) to perform subsequent amplification reaction; The heating module (419) is moved left and right to a position aligned with the amplification chamber (301j), and the heating module (419) is moved upward. The heating module (419) is attached to the lower end of the corresponding amplification chamber (301j) through the heating cavity (419a). The heating element in the heating module (419) is activated to heat the amplification chamber (301j) to a set temperature. The polymerase gradually adds deoxyribonucleoside triphosphates along the 3' end of the template nucleic acid to synthesize a new nucleic acid chain. When the polymerase synthesizes along the template chain to the probe binding position, the probe is cut by its 5'-3' exonuclease activity, so that the fluorescent reporter group is separated from the quenching group, and the fluorescent signal is released, so that the heating cavity ( 419a) leaves the amplification chamber (301j), completing one cycle, the detection probe (418) moves to the same level as the amplification chamber (301j), and according to the fluorescent groups of different wavelengths connected to the TapMan probe, the detection probe (418) is rotated so that the excitation light source of the corresponding wavelength on the detection probe (418) and the detection probe (418) are aligned with the amplification chamber (301j), and the fluorescence intensity of different amplification chambers (301j) is detected to complete one cycle; return to the above detection step until the set number of cycles is reached, and the signal detection is completed.

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