WO2025103413A1 - High-throughput full-automatic nucleic acid detection instrument and method - Google Patents

Abstract

A high-throughput full-automatic nucleic acid detection instrument and method. The high-throughput full-automatic nucleic acid detection instrument comprises: a base; a transfer module, which is arranged on the base; a pipetting module, which is used for adding detection samples into a plurality of plate holes in a nucleic acid extraction plate on the transfer module; a nucleic acid extraction module, which comprises a plurality of magnetic head assemblies, wherein the magnetic head assembly comprises a driving device, a sleeve, a magnetic rod and magnetic beads, the driving device being used for driving the magnetic rod to be extended into or withdrawn from the sleeve, the magnetic rod being used for attracting or releasing the magnetic beads, when the magnetic rod is extended into the sleeve, the magnetic beads being attracted to an outer wall of the sleeve, and the magnetic beads being transferred between the plate holes along with the sleeve, and when the magnetic rod is withdrawn from the sleeve, the magnetic beads with nucleic acids adsorbed being released in the plate holes, so as to transfer the nucleic acids; and a PCR detection module, which is used for performing PCR detection on a PCR detection plate added with the nucleic acids. Regional division and pressure control are performed on modules on the basis of a nucleic acid detection process, and contamination leakage of a PCR detection region is prevented and controlled by means of a closable chamber door and a negative pressure system.

Description

High-throughput fully automatic nucleic acid detection instrument and method

CROSS-REFERENCE TO RELATED APPLICATIONS

This application claims priority to Chinese patent application number 202311535375.3, filed on November 17, 2023, and entitled “High-throughput fully-automatic nucleic acid detection instrument and method”, the entire text of which is hereby incorporated by reference.

Technical Field

The present application relates to the field of nucleic acid detection technology, and specifically to a high-throughput fully automatic nucleic acid detection instrument and method.

Background Art

With the development of nucleic acid detection technology, nucleic acid detection has been widely used in various fields such as pathogen diagnosis, scientific research, environmental monitoring, etc. Especially in recent years, with the promotion of nucleic acid detection technology in the field of blood safety, fluorescent PCR technology has been widely used.

Currently, the most widely used nucleic acid detection instruments are semi-automatic instruments, which are divided into nucleic acid extractors and fluorescent PCR instruments. The two are used together, and cannot automatically seal, transfer, and analyze the test results of PCR reaction tubes. They need to be performed manually and multiple devices need to be operated. There are also a small number of fully automatic nucleic acid detection instruments that simply connect nucleic acid extraction and PCR amplification detection together. However, the detection throughput and detection speed cannot meet the needs of large sample volumes and rapid detection. At the same time, sharing a set of pipette needles for sample addition, nucleic acid extraction, and PCR reaction solution preparation will cause cross contamination of sample detection.

The main limiting factors that make the existing technical solutions unable to achieve rapid detection of large sample volumes and prone to cross contamination are: the nucleic acid extraction link is a key link, which takes a long time and is prone to cross contamination. Although the existing solutions have achieved the automation of nucleic acid extraction, the existing implementation methods mostly use pipetting devices or oscillating devices to simulate manual operations for nucleic acid extraction. There are mainly two methods. Existing method 1: Nucleic acid extraction uses a pipette to repeatedly blow and suck the sample and nucleic acid extraction reagent to mix, and uses a magnetic module to separate magnetic beads outside the container such as a nucleic acid extraction tube or plate, and then uses a pipette to suck and discard each nucleic acid extraction solution. The defect of this method is that the use of a sample needle and a pipette to repeatedly suck and discard waste liquid is prone to aerosol contamination, resulting in false positive test results. At the same time, because a limited number of pipette needles are used for multiple times to suck and discard waste liquid, the time required is long and the extraction speed is slow. In addition, the residue of the extract on the tube wall affects the purity of nucleic acid extraction. Existing method 2: Use a temperature-controlled oscillation module to openly oscillate and mix the extraction plate, which is easy to cause liquid splashing and aerosol contamination. In addition, this method also uses a magnetic module outside the nucleic acid extraction tube or plate container to separate the magnetic beads, and then uses a pipette to aspirate and discard each nucleic acid extraction solution. The disadvantage of this method is that using a sample needle and a pipette to aspirate and discard waste liquid multiple times is prone to aerosol contamination, resulting in false positive test results. The limited number of pipette needles used for multiple extractions takes a long time and the extraction speed is slow. In addition, the extraction solution remains on the tube wall, affecting the purity of the nucleic acid extraction.

Summary of the invention

The present application aims to provide a high-throughput fully automatic nucleic acid detection instrument and method, which aims to solve the problems in the prior art of using a sample needle and a pipette tip to repeatedly blow and aspirate samples to mix and aspirate waste liquid, which easily generates aerosol contamination, and the long time required to aspirate waste liquid, slow extraction speed, and residual extract on the tube wall that affects the purity of nucleic acid extraction.

On the one hand, the present application provides a high-throughput fully automatic nucleic acid detection instrument, the high-throughput fully automatic nucleic acid detection instrument comprising:

Base;

A transfer module, the transfer module is arranged on the base, the transfer module is used to carry a nucleic acid extraction plate, a nucleic acid release plate and a PCR detection plate, wherein the nucleic acid extraction plate, the nucleic acid release plate and the PCR detection plate are all provided with a plurality of plate holes, the plate holes of the nucleic acid extraction plate and the nucleic acid release plate are used to accommodate extraction reagents, and the plate holes of the PCR detection plate are used to accommodate PCR reaction liquid;

A mechanical gripper, wherein the mechanical gripper is used to transport the nucleic acid extraction plate, the nucleic acid release plate and the PCR detection plate;

A pipetting module, which is used to add the test sample to a plurality of plate wells on the nucleic acid extraction plate on the transfer module;

A nucleic acid extraction module, wherein the nucleic acid extraction module comprises a plurality of magnetic head assemblies, wherein the magnetic head assemblies comprise a driving device, a sleeve, a magnetic rod and magnetic beads, wherein the driving device is used to drive the magnetic rod to extend into or out of the sleeve, wherein the magnetic rod is used to adsorb or release the magnetic beads, wherein the magnetic beads are used to adsorb nucleic acids, wherein when the magnetic rod is extended into the sleeve, the magnetic beads are adsorbed onto the outer wall of the sleeve, and the magnetic beads follow the sleeve to transfer between the plate holes; when the magnetic rod is pulled out of the sleeve, the magnetic beads adsorbed with nucleic acids are released into the plate holes, thereby transferring the nucleic acids;

A PCR detection module is disposed on the base, and is used to perform PCR detection on the PCR detection plate to which nucleic acid is added to obtain a detection result.

Optionally, the number of the mechanical grippers may be one or more. In a preferred embodiment of the present application, the mechanical grippers include a first mechanical gripper and a second mechanical gripper, wherein the first mechanical gripper is used to transport the nucleic acid extraction plate and the nucleic acid release plate, and the second mechanical gripper is used to transport the PCR detection plate.

In some embodiments, the driving device drives the sleeve to rotate and stir the solution in the plate hole, and the outer wall of the sleeve is provided with corrugations.

In the present application, by providing corrugations on the outer wall of the sleeve, on the one hand, splashing of liquid during the stirring process can be prevented; On the other hand, it can make the small volume of solution concentrated at the bottom of the tube stirred more evenly.

In some embodiments, the base includes a nucleic acid extraction area, a PCR detection area and an isolation door, the isolation door is arranged between the nucleic acid extraction area and the PCR detection area, the pipetting module and the nucleic acid extraction module are located in the nucleic acid extraction area, and the PCR detection module is located in the PCR detection area.

In some embodiments, a sealing module is provided on the base, and the sealing module is arranged in the PCR detection area. The sealing module is used to seal the PCR detection plate added with nucleic acid transported by the mechanical gripper; the PCR detection module is used to detect the PCR detection plate after sealing.

In some embodiments, a transmission module is provided between the PCR detection module and the nucleic acid extraction module, and the transmission module passes through the isolation door. The isolation door cuts off the transmission module when it is closed, and when the isolation door is opened, the transmission module moves the PCR detection plate added with nucleic acid from the nucleic acid extraction module to the PCR detection module.

In some embodiments, a protective rear plate and a protective side plate are vertically arranged on the edge of the base, and the protective rear plate and the protective side plate surround various devices on the base.

In some embodiments, the mechanical gripper and the pipetting module are arranged on the protective back plate, and the base is provided with a barcode scanning module, a sample module, a consumables carrying position and a reagent module. The sample module is located below the pipetting module, and the sample module is used to carry the test sample. The barcode scanning module is used to perform barcode scanning on the test samples, reagents and consumables in the sample module, the transfer module, the reagent module and the consumables carrying position.

In some embodiments, a waste conveying channel is provided on the base, and the waste conveying channel is located between the transfer module and the sample module. A waste collection module is provided at one end of the waste conveying channel, and a spare consumables position is provided on the protective rear plate.

In some embodiments, an air filter cover is provided on the top of the protective rear plate and the protective side plate, an exhaust fan and an air filter are installed on the air filter cover, and an ultraviolet lamp is provided in the air filter cover, and the ultraviolet lamp is used for disinfection.

In the present application, the base is configured as a layer or includes an upper layer and a lower layer, and a transfer channel is provided between the upper layer and the lower layer, and the transfer channel is used to transfer items; the base shares one mechanical gripper, or corresponding mechanical grippers are provided in different areas of the base.

In the present application, each functional module can be arranged on the upper layer or the lower layer of the base according to actual needs. As an embodiment of the present application, the transfer module, the liquid transfer module and the nucleic acid extraction module of the high-throughput fully automatic nucleic acid detection instrument of the present application are arranged on the upper layer of the base, and the PCR detection module and the sealing module are arranged on the lower layer of the base.

On the other hand, the present application provides a high-throughput fully automatic nucleic acid detection method, the high-throughput fully automatic nucleic acid detection method Methods, including:

The pipetting module adds the test sample into a plurality of plate wells on the nucleic acid extraction plate on the transfer module;

The mechanical gripper grabs the nucleic acid extraction plate and places it into the nucleic acid extraction module;

The plurality of sleeves on the nucleic acid extraction module extend into the plurality of plate holes on the nucleic acid extraction plate for stirring;

One or more magnetic bars of the nucleic acid extraction module extend into the sleeve, so that the magnetic beads adsorbed with nucleic acid are adsorbed on the outer wall of the sleeve;

The plurality of magnetic head assemblies of the nucleic acid extraction module are separated from the nucleic acid extraction plate as a whole;

The multiple magnetic head assemblies of the nucleic acid extraction module are integrally extended into the nucleic acid release plate, and the magnetic rod is pulled out of the sleeve to release the magnetic beads into the multiple plate holes of the nucleic acid release plate;

The plurality of sleeves on the nucleic acid extraction module are rotated and stirred in the plurality of plate holes on the nucleic acid release plate, so that the nucleic acid on the magnetic beads is eluted onto the nucleic acid release plate;

One or more magnetic bars of the nucleic acid extraction module extend into the sleeve, so that the magnetic beads are adsorbed on the outer wall of the sleeve;

The mechanical gripper moves the nucleic acid release plate with nucleic acid to the transfer module;

The liquid transfer module adds the solution in the nucleic acid release plate and the PCR reaction solution into a plurality of plate wells on the PCR detection plate on the transfer module to obtain the PCR detection plate to which nucleic acid is added;

The mechanical gripper transports the PCR detection plate with added nucleic acid to the PCR detection module for detection to obtain the detection result.

In the present application, the nucleic acid extraction reagent can be pre-stored in the plate wells, or added to the plate wells through a pipetting module.

The present application provides a high-throughput fully automatic nucleic acid detection instrument, which realizes the rapid transfer of magnetic beads. The magnetic bead transfer process does not use the method of transferring waste liquid by aspirating with a sample needle. The waste liquid in each step is retained in the plate well, avoiding cross contamination during the transfer process. In this way, the defects of the existing method of using a sample needle and a pipette tip to repeatedly blow and aspirate samples to mix and aspirate waste liquid, which easily generates aerosol pollution, and the long time required for aspirating waste liquid, slow extraction speed, and residual extract on the tube wall affecting the purity of nucleic acid extraction are avoided. At the same time, the technical solution of the present application also overcomes the defects of aerosol pollution caused by the open agitation and mixing of the oscillation module in the prior art.

In addition, the high-throughput fully automatic nucleic acid detection instrument of the present application separates the nucleic acid extraction area and the PCR detection area by setting an isolation door. The PCR detection area is designed with negative pressure, and the exhaust speed and direction of the exhaust fan realize a decreasing pressure gradient, which conforms to the reasonable pressure difference setting of nucleic acid detection, prevents contamination during the detection process, and cooperates with the air filter to further prevent external dust from interfering with the detection. The waste collection module of the high-throughput fully automatic nucleic acid detection instrument in the present application is set on the outside of the base and is protected by a metal shell on the outer layer. It is transported through the waste conveying channel. The waste consumables are transported after use, reducing the contamination caused by the waste collection module being exposed or close to the nucleic acid extraction area and the PCR detection area. The technical solution of the present application significantly improves the accuracy of the test results and reduces the risk of contamination through the coordination of the above structure.

In addition, the high-throughput fully automatic nucleic acid detection instrument of the present application has a large sample throughput, up to 576 samples can be loaded at one time, and it supports multiple rounds of continuous loading, with a detection throughput of up to 4608 samples, which is a huge improvement over the 96 sample positions of similar devices, and the detection speed is also faster. The detection sensitivity of the high-throughput fully automatic nucleic acid detection instrument of the present application is also higher, up to 40IU/mL. For some samples, the detection sensitivity can even be as low as 3IU/mL, which is much higher than similar instruments already on the market.

BRIEF DESCRIPTION OF THE DRAWINGS

In order to more clearly illustrate the technical solutions in the embodiments of the present application, the drawings required for use in the description of the embodiments will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present application. For those skilled in the art, other drawings can be obtained based on these drawings without creative work.

FIG1 is a schematic diagram of the structure of a high-throughput fully automatic nucleic acid detection instrument provided in an embodiment of the present application;

FIG2 is a schematic diagram of the internal structure of a high-throughput fully automatic nucleic acid detection instrument provided in an embodiment of the present application;

FIG3 is a schematic diagram of cannula stirring in a high-throughput fully automatic nucleic acid detection instrument provided in an embodiment of the present application;

4 is a schematic diagram of a high-throughput fully-automatic nucleic acid detection instrument provided in an embodiment of the present application, in which a magnetic rod is inserted into a cannula and magnetic beads are adsorbed and collected under the cannula;

FIG5 is a schematic diagram of the entire magnetic head assembly being separated from the plate hole in the high-throughput fully-automatic nucleic acid detection instrument provided in an embodiment of the present application;

6 is a schematic diagram of the release of magnetic beads after the magnetic rod of the magnetic head assembly in the high-throughput fully-automatic nucleic acid detection instrument provided in an embodiment of the present application is separated from the casing;

FIG7 is a schematic diagram of a cannula in a high-throughput fully-automatic nucleic acid detection instrument provided in an embodiment of the present application;

FIG8 is a schematic diagram of the structure of a high-throughput fully-automatic nucleic acid detection instrument provided in an embodiment of the present application; and

FIG9 is a schematic flow chart of an embodiment of a high-throughput fully-automatic nucleic acid detection method provided in an embodiment of the present application.

In the figure: 1-barcode scanning module; 2-sample module; 3-pipette module; 4-transfer module; 5-reagent module; 6-consumables carrying position; 7-nucleic acid extraction module; 8-transmission module; 9-isolation door; 10-sealing module; 11-PCR detection module; 12-first mechanical gripper; 13-second mechanical gripper; 14-waste conveying channel; 141-waste collection module; 15-spare consumables position; 16-air filter cover; 17-protective side panel; 18-protective rear panel; 19-exhaust fan; 20-base; 30-magnetic head assembly; 31-sleeve; 32-corrugation; 33-magnetic beads; 34-plate hole; 35-magnetic rod; 36-transfer channel.

DETAILED DESCRIPTION

The following will be combined with the drawings in the embodiments of the present application to clearly and completely describe the technical solutions in the embodiments of the present application. Obviously, the described embodiments are only part of the embodiments of the present application, not all of the embodiments. Based on the embodiments in the present application, all other embodiments obtained by those skilled in the art without creative work are within the scope of protection of this application.

In the description of the present application, it should be understood that the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "up", "down", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inside", "outside" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the accompanying drawings, which are only for the convenience of describing the present application and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, be constructed and operated in a specific orientation, and therefore cannot be understood as a limitation on the present application. In addition, the terms "first" and "second" are only used for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Therefore, the features defined as "first" and "second" may explicitly or implicitly include one or more features. In the description of the present application, the meaning of "multiple" is two or more, unless otherwise clearly and specifically defined.

In this application, the word "exemplary" is used to mean "used as an example, illustration, or description." Any embodiment described in this application as "exemplary" is not necessarily to be construed as being preferred or advantageous over other embodiments. The following description is given to enable any technician in the field to implement and use the present application. In the following description, details are listed for the purpose of explanation. It should be understood that a person of ordinary skill in the art can recognize that the present application can be implemented without using these specific details. In other instances, well-known structures and processes will not be elaborated in detail to avoid obscuring the description of the present application with unnecessary details. Therefore, the present application is not intended to be limited to the embodiments shown, but is consistent with the widest scope consistent with the principles and features disclosed in the present application.

The embodiments of the present application provide a high-throughput fully-automatic nucleic acid detection instrument and method, which are described in detail below.

First, a high-throughput fully-automatic nucleic acid detection instrument is provided in an embodiment of the present application, and the high-throughput fully-automatic nucleic acid detection instrument includes: a base; a transfer module, the transfer module is arranged on the base, the transfer module is used to carry a nucleic acid extraction plate, a nucleic acid release plate and a PCR detection plate, wherein the nucleic acid extraction plate, the nucleic acid release plate and the PCR detection plate are each provided with a plurality of plate holes, the plate holes of the nucleic acid release plate are used to accommodate the extracted nucleic acid, and the plate holes of the PCR detection plate are used to accommodate the PCR reaction solution; a first mechanical gripper, the first mechanical gripper is used to transport the nucleic acid extraction plate; a second mechanical gripper, the second mechanical gripper is used to transport the PCR detection plate; a pipetting module, the pipetting module is used to add the detection sample to the plurality of plate holes on the nucleic acid extraction plate on the transfer module; a nucleic acid extraction module, the nucleic acid extraction module includes a plurality of magnetic head assemblies, the magnetic head assembly includes a driving device, a sleeve, a magnetic rod and magnetic beads, the driving device is used to drive the magnetic rod to extend into or out of the sleeve, the magnetic rod is used to adsorb or release the magnetic beads, the magnetic beads are used to adsorb the nucleic acid, when the magnetic rod is extended into the sleeve, the magnetic beads are adsorbed on the outer wall of the sleeve, and the magnetic beads follow the sleeve in the plate holes transfer between; when the magnetic rod is pulled out of the sleeve, the magnetic beads adsorbed with nucleic acid are released in the plate holes, thereby transferring the nucleic acid; PCR detection module, the PCR detection module is arranged on the base, and the PCR detection module is used to perform PCR detection on the PCR detection plate added with nucleic acid to obtain the detection result.

Referring to Figures 1 to 7, in the embodiments of the present application, the high-throughput fully automatic nucleic acid detection instrument includes: a base 20, a transfer module 4, a first mechanical gripper 12, a second mechanical gripper 13, a pipetting module 3, a nucleic acid extraction module 7, and a PCR detection module 11. In other embodiments, the position of each functional module can be changed according to the specific arrangement form and space requirements, and these simple changes all fall within the scope of protection of the present invention.

The base 20 is a platform.

In the embodiment of the present application, the transfer module 4 is disposed on the base 20, and the transfer module 4 is used to carry the nucleic acid extraction plate, the nucleic acid release plate, and the PCR detection plate. The nucleic acid extraction plate, the nucleic acid release plate, and the PCR detection plate are all provided with a plurality of plate holes 34, the plate holes 34 of the nucleic acid release plate are used to accommodate the extraction reagent, and the plate holes 34 of the PCR detection plate are used to accommodate the PCR reaction solution.

In the embodiment of the present application, the first mechanical gripper 12 is used to transport the nucleic acid extraction plate and the nucleic acid release plate.

In the embodiment of the present application, the second mechanical gripper 13 is used to transport the PCR detection plate.

In the embodiment of the present application, the pipetting module 3 is used to add the test sample to the multiple plate wells 34 on the nucleic acid extraction plate on the transfer module 4. Specifically, 96 plate wells 34 are provided on the nucleic acid extraction plate, the nucleic acid release plate and the PCR detection plate.

In the embodiment of the present application, the nucleic acid extraction module 7 includes multiple magnetic head assemblies 30, and the magnetic head assembly 30 includes a driving device, a sleeve 31, a magnetic rod 35 and magnetic beads 33. The driving device is used to drive the magnetic rod 35 to extend in and out of the sleeve 31. The magnetic rod 35 is used to adsorb or release the magnetic beads 33. The magnetic beads 33 are used to adsorb nucleic acids. When the magnetic rod 35 is extended into the sleeve 31, the magnetic beads 33 are adsorbed on the outer wall of the sleeve 31, and the magnetic beads 33 follow the sleeve 31 to transfer between the plate holes 34; when the magnetic rod 35 is pulled out of the sleeve 31, the magnetic beads 33 adsorbed with nucleic acids are released in the plate holes 34, thereby transferring the nucleic acids.

In the embodiment of the present application, the PCR detection module 11 is disposed on the base 20, and the PCR detection module 11 is used to perform PCR detection on the PCR detection plate added with nucleic acid to obtain the detection result.

The present application uses a magnetic bar 35 to transfer magnetic beads 33 for nucleic acid extraction. The magnetic head assembly 30 with the magnetic bar 35 can move up and down, and the sleeve 31 rotates in the plate hole 34 to mix the sample and the nucleic acid extraction reagents. When the magnetic bar 35 is inserted into the sleeve 31 and extends into the plate hole 34, the magnetic beads 33 are adsorbed and gathered on the sleeve 31 outside the magnetic bar 35 by the magnetic field of the magnetic bar 35 and separated from the extraction solution. After the sleeve 31 adsorbs the magnetic beads 33, the magnetic bar 35 is lifted upward from the plate hole 34, and the magnetic beads 33 are released into the next plate hole 34. The magnetic head assembly 30 can transfer the magnetic beads 33 of 96 plate holes 34 at one time, realizing the rapid transfer of the magnetic beads 33. The transfer process of the magnetic beads 33 does not use the method of aspirating and discarding the waste liquid by using a sample needle. The waste liquid of each step is retained in the plate hole 34 to avoid cross contamination during the transfer process. The present invention overcomes the problems in the prior art that the sample is repeatedly blown and sucked by the sample needle and the pipette tip to mix and suck the waste liquid, which easily generates aerosol pollution, takes a long time to suck the waste liquid, has a slow extraction speed, and the extract is stuck on the tube wall. Residual defects that affect the purity of nucleic acid extraction.

In the embodiment of the present application, the driving device drives the sleeve 31 to rotate and stir the solution in the plate hole 34, and the outer wall of the sleeve 31 is provided with corrugations 32. In the new plate hole 34, the magnetic rod 35 is pulled upward from the sleeve 31, and the magnetic rod 35 is separated from the magnetic adsorption of the magnetic beads 33, and the sleeve 31 is rotated in the extraction reagent to achieve the release of the magnetic beads 33 in the extraction solution. The sleeve 31 has corrugations 32, which can better play the effect of stirring the sample, magnetic beads 33 and extraction reagent. In this way, the present application overcomes the defects of aerosol contamination caused by repeated blowing and suction mixing with a sample adding needle and open oscillation mixing with an oscillation module in the existing method.

In an embodiment of the present application, the base 20 includes a nucleic acid extraction area, a PCR detection area and an isolation door 9, the isolation door 9 is arranged between the nucleic acid extraction area and the PCR detection area, the pipetting module 3 and the nucleic acid extraction module 7 are located in the nucleic acid extraction area, and the PCR detection module 11 is located in the PCR detection area.

In the embodiment of the present application, a sealing module 10 is provided on the base 20, and the sealing module 10 is arranged in the PCR detection area. The sealing module 10 is used to seal the PCR detection plate added with nucleic acid transported by the second mechanical gripper 13; the PCR detection module 11 is used to detect the PCR detection plate after sealing.

In an embodiment of the present application, a transmission module 8 is provided between the PCR detection module 11 and the nucleic acid extraction module 7. The transmission module 8 passes through an isolation door 9. The isolation door 9 cuts off the transmission module 8 when it is closed. When the isolation door 9 is opened, the transmission module 8 moves the PCR detection plate added with nucleic acid from the nucleic acid extraction module 7 to the sealing module 10 or the PCR detection module 11.

In the embodiment of the present application, a protective rear plate 18 and a protective side plate 17 are vertically arranged on the edge of the base 20 , and the protective rear plate 18 and the protective side plate 17 surround various components on the base 20 .

In the embodiment of the present application, the first mechanical gripper 12, the second mechanical gripper 13 and the pipetting module 3 are arranged on the protective rear plate 18, and the base 20 is provided with a barcode scanning module 1, a sample module 2, a consumables carrying position 6 and a reagent module 5. The sample module 2 is located below the pipetting module 3, and the sample module 2 is used to carry the test sample. The barcode scanning module 1 is used to perform barcode scanning on the test sample, reagents and consumables in the sample module 2, the transfer module 4, the reagent module 5 and the consumables carrying position 6.

In the embodiment of the present application, an air filter cover 16 is provided on the top of the protective rear plate 18 and the protective side plate 17, and an exhaust fan 19 and an air filter are installed on the air filter cover 16. An ultraviolet lamp is provided in the air filter cover 16, and the ultraviolet lamp is used for disinfection.

The high-throughput fully automatic nucleic acid detection instrument in this application is divided into a nucleic acid extraction area and a PCR detection area, and the PCR detection area is separated independently, and the PCR detection area is isolated by an independent chassis. The PCR detection area is negative pressure, and the PCR detection plate is transmitted through an openable and closable isolation door 9 to prevent the leakage of amplification products from causing pollution in other areas. The air filter cover 16 and the PCR detection area are negative pressure designs, which play a role in preventing external dust from interfering with the detection. An exhaust fan 19 and an air filter are provided, and the air pressure gradient of the nucleic acid extraction area and the PCR detection area is formed by the exhaust speed and direction of the fan to realize the air pressure gradient of decreasing air pressure, which meets the reasonable pressure difference setting of nucleic acid detection and prevents pollution during the detection process. An ultraviolet lamp is installed in the air filter cover 16, and the ultraviolet lamp irradiates and disinfects each detection area after the detection is completed to eliminate nucleic acid contamination in the detection. The invention solves the defects of the prior art that the functional areas are not effectively divided, the air flow is not effectively controlled, and the detection cross contamination is easily caused.

In the embodiment of the present application, a waste conveying channel 14 is provided on the base 20, and the waste conveying channel 14 is located between the transfer module 4 and the sample module 2. A waste collection module 141 is provided at one end of the waste conveying channel 14, and the waste collection module 141 is located outside the base 20. A spare consumable position 15 is provided on the protective back plate 18. The present application moves waste to the waste collection module 141 through the waste conveying channel 14. The waste collection module 141 is arranged outside the base 20, outside the nucleic acid extraction area and the PCR detection area, and has a small opening, which is only used for discarding pipette tips. The inner layer of the waste collection module 141 is a plastic garbage bag, and the outer layer is protected by a metal shell. The defect of the prior art that the waste garbage bin is open and is close to the liquid preparation area in the detection area, which is easy to cause waste pollution, is solved.

In the embodiment of the present application, the pipetting module 3 may contain 1 to 96 sample injection channels, and a sample injection needle or a pipette tip may be used. The sample positions in the sample module 2 may be 1 to 1000. The transfer module 4 and the reagent module 5 may be 1 to 100. The sealing module 10 may adopt a heat sealing film module or an adhesive film sealing film module. The transmission module 8 may be placed in the PCR detection area or in other areas.

The PCR detection plate entry and exit port of the PCR detection module 11 in the present application can be opened and closed by a push-pull method or by a flip-cover method.

As shown in Fig. 7, the upper part of the sleeve 31 is provided with a corrugated structure 32 to prevent the liquid from splashing out of the plate hole 34 during stirring. The middle and lower parts of the sleeve 31 have a corrugated structure 32 to make the solution, especially the small volume solution concentrated at the bottom of the tube, stirred more evenly.

Refer to Figure 9, which is a schematic diagram of an embodiment of a high-throughput fully-automatic nucleic acid detection method provided in an embodiment of the present application. The high-throughput fully-automatic nucleic acid detection method includes 201-212:

201 . The pipetting module 3 adds the test sample into the multiple plate wells 34 on the nucleic acid extraction plate on the transfer module 4 .

In the embodiment of the present application, before step 201, the barcode scanning module 1 can perform barcode scanning on the test samples, reagents and consumables in the sample module 2, the consumables holding position 6 and the reagent module 5. The scanning information is recorded in the instrument computer, thereby realizing barcode monitoring of the entire detection process.

In the embodiment of the present application, the pipetting module 3 includes a robotic arm and a sample adding needle or pipette tip, which can insert the pipette tip in the consumable module, and then pipette the sample in the sample module 2 and the nucleic acid reagent in the reagent module 5, and add them to the multiple plate holes 34 in each nucleic acid extraction plate in the transfer module 4.

202. Control the first mechanical gripper 12 to grip the nucleic acid extraction plate and place it on the nucleic acid extraction module 7 to perform nucleic acid extraction.

In the embodiment of the present application, after the pipetting is completed, the first mechanical gripper 12 is controlled to grab the nucleic acid extraction plate and place it on the nucleic acid extraction module 7 for nucleic acid extraction.

203. The multiple sleeves 31 on the nucleic acid extraction module 7 extend into the multiple plate holes 34 on the nucleic acid extraction plate to rotate and stir.

In the embodiment of the present application, the sleeves 31 of the multiple magnetic head assemblies 30 correspond to the plate holes 34 on the multiple nucleic acid extraction plates. As shown in FIG3 , the multiple sleeves 31 on the nucleic acid extraction module 7 extend into the multiple plate holes 34 on the nucleic acid extraction plate for stirring. The sleeves 31 can be moved up and down and rotated to perform rotational stirring, so that the sample and the nucleic acid extraction reagents are mixed.

204 . The multiple magnetic bars 35 of the nucleic acid extraction module 7 extend into the sleeve 31 , so that the magnetic beads 33 adsorbed with nucleic acid are adsorbed on the outer wall of the sleeve 31 .

As shown in FIG. 4 , when the magnetic rod 35 extends into the sleeve 31 , the magnetic beads 33 are adsorbed on the outer wall of the sleeve 31 , and the magnetic beads 33 are collected by the sleeve 31 . The magnetic beads 33 follow the sleeve 31 and transfer between the plate holes 34 .

205. The multiple magnetic head assemblies 30 of the nucleic acid extraction module 7 are completely separated from the nucleic acid extraction plate.

As shown in FIG. 5 , the multiple magnetic head assemblies 30 of the nucleic acid extraction module 7 are completely separated from the nucleic acid extraction plate.

206. The multiple magnetic head assemblies 30 of the nucleic acid extraction module 7 are integrally extended into the nucleic acid release plate, and the magnetic rod 35 is pulled out of the sleeve 31 to release the magnetic beads 33 to enter the multiple plate holes 34 of the nucleic acid release plate.

As shown in FIG. 6 , the multiple magnetic head assemblies 30 of the nucleic acid extraction module 7 are integrally extended into the nucleic acid release plate, and the magnetic rod 35 is pulled out of the sleeve 31 to release the magnetic beads 33 into the multiple plate holes 34 of the nucleic acid release plate, thereby realizing the transfer of the magnetic beads 33 .

207. The multiple sleeves 31 on the nucleic acid extraction module 7 are rotated and stirred in the multiple plate holes 34 on the nucleic acid release plate, so that the nucleic acid on the magnetic beads 33 is eluted onto the nucleic acid release plate.

The sleeve 31 rotates in the extraction reagent in the nucleic acid release plate to release the magnetic beads 33 in the extraction solution. The sleeve 31 has ripples 32, which can better stir the sample, magnetic beads 33 and extraction reagent. The nucleic acid is retained in the nucleic acid release plate through the extraction of the extraction reagent.

208 . The multiple magnetic bars 35 of the nucleic acid extraction module 7 extend into the sleeve 31 , so that the magnetic beads 33 are adsorbed on the outer wall of the sleeve 31 .

In the embodiment of the present application, after the sample is lysed by the nucleic acid extraction reagent, the nucleic acid is adsorbed by the magnetic beads 33, and washed, the magnetic beads 33 adsorbed with nucleic acid are transferred from the nucleic acid extraction module 7 to the nucleic acid release plate filled with elution solution for elution, and the magnetic beads 33 are collected and separated by the magnetic rod 35 and the sleeve 31 to complete the extraction of nucleic acid.

209 . The first mechanical gripper 12 moves the nucleic acid release plate with nucleic acid to the transfer module 4 .

The first mechanical gripper 12 transfers the nucleic acid release plate from which nucleic acid extraction has been completed to the transfer module 4 .

210. The pipetting module 3 adds the solution in the nucleic acid release plate and the PCR reaction solution into the multiple plate wells 34 on the PCR detection plate on the transfer module 4 to obtain a PCR detection plate with added nucleic acid.

In the embodiment of the present application, the sample adding needle of the pipette module 3 uses the pipette head in the consumable module to absorb the PCR reaction reagents in the reagent module 5 to prepare the PCR reaction solution, and then absorbs and distributes the PCR reaction solution into the plate hole 34 of the PCR detection plate of the transfer module 4. Then, the solution in the nucleic acid release plate is transferred to the plate hole 34 of the PCR detection plate of the transfer module 4, thereby completing the Preparation of PCR reaction solution.

211. The second mechanical gripper 12 transfers the PCR detection plate with added nucleic acid to the transmission module 8 for sealing, thereby obtaining the sealed PCR detection plate.

In the embodiment of the present application, the first mechanical gripper 12 moves the PCR test plate with added nucleic acid from the transfer module 4 to the transmission module 8. The isolation door 9 is opened, and the transmission module 8 transports the PCR test plate with added nucleic acid to the PCR detection area through the transmission track, and then the isolation door 9 is closed. The second mechanical gripper 13 transfers the PCR test plate to the sealing module 10, and the PCR test plate in the sealing module 10 is sealed.

212. The second mechanical gripper 13 transfers the sealed PCR detection plate to the PCR detection module 11 for detection to obtain the detection result.

The second mechanical gripper 13 transfers the sealed PCR detection plate to the PCR detection module 11 for fluorescent PCR amplification detection. After the detection is completed, the software automatically analyzes the detection data and generates the detection results.

The discarded pipette tips in the detection process are stored in the waste collection module 141, and the various extraction waste liquids in the extraction process are left in the corresponding extraction plate, without the need to collect waste liquid separately, and waste disposal is convenient. The protective rear plate 18 and the protective side plate 17 are in a closed state and locked during the detection process, ensuring the airtightness and personnel safety of the detection process. The air filter cover 16 plays a role in preventing external dust from interfering with the detection, and the air filter cover 16 is equipped with an exhaust fan 19 and an air filter. The exhaust speed and direction of the exhaust fan 19 realize the air pressure gradient of the sample area, the nucleic acid extraction area and the PCR detection area to decrease the air pressure, which meets the reasonable pressure difference setting of nucleic acid detection and prevents pollution during the detection process. At the same time, the PCR detection area is isolated by an independent chassis, and the chassis is negative pressure. The PCR detection plate is transmitted through the openable and closable isolation door 9 to prevent the leakage of the amplification product from causing pollution in other areas. An ultraviolet lamp is installed in the air filter cover 16. After the detection is completed, the ultraviolet lamp irradiates each detection area to eliminate nucleic acid contamination in the detection.

The high-throughput fully automatic nucleic acid detection instrument of the present application has a large sample throughput. The sample module 2 can hold 1 to multiple rows of samples, with up to 576 samples loaded at one time. It also supports multiple rounds of continuous loading, and can take 1, 6, 8, 24, and 48 samples mixed into 1 hole for combined detection. The detection throughput of each experiment is as high as 4608 samples, which is a huge improvement over the 96 sample positions of similar devices currently available. The nucleic acid extraction module 7 of the high-throughput fully automatic nucleic acid detection instrument of the present application contains 48/96 magnetic head assemblies 30, which can extract 96-hole samples at a time, which is 12 times faster than the 1 to 8-pin pipetting device used in the existing device to extract 1-8-hole samples at a time. The extraction and transfer magnetic beads 33 process does not use the method of aspirating and discarding waste liquid by using a sample needle, which also saves extraction time. The total detection time of 558 plasma samples and 3 quality control products from blood donors using the instrument of the present application in combination with the hepatitis B virus/hepatitis C virus/human immunodeficiency virus (1+2) type nucleic acid detection reagent is 4 hours. It is 2.5 hours faster than the conventional instrument Roche cobas AmpliPrep/cobas TaqMan detection system, which takes 6.5 hours to detect similar samples. The entire detection time for testing 4464 plasma samples and 3 quality control products from blood donors is 8.25 hours. The example verification is as follows:

(1) Example of mixed testing of 6 samples

The instrument of this application was used with the hepatitis B virus/hepatitis C virus/human immunodeficiency virus (1+2) nucleic acid detection reagent (purchased from Beijing Wantai Biopharmaceutical Co., Ltd.) to test 558 plasma samples and 3 quality control samples from blood donors. The test time was calculated.

Step 1: Complete the barcode scanning of 3 quality control samples and 558 samples, and pool and mix every 6 plasma samples into 1 sample. The time to complete the sample loading is 45 minutes;

Step 2: Reagent addition and nucleic acid extraction time is 96 minutes;

Step 3: PCR reagent preparation time is 6 minutes;

Step 4: The PCR reaction system construction time is 13 minutes;

Step 5: PCR sealing and PCR detection time is 80 minutes.

The total detection time is 240 minutes, or 4 hours, which is 2.5 hours faster than the conventional instrument Roche cobas AmpliPrep/cobas TaqMan detection system, which takes 6.5 hours to detect similar samples.

(2) Example of mixed testing of 48 samples

The instrument of this application was used with the hepatitis B virus/hepatitis C virus/human immunodeficiency virus (1+2) nucleic acid detection reagent to detect 4464 plasma samples and 3 quality control samples from blood donors. The test time was calculated.

Step 1: Complete the barcode scanning of 3 quality control products and 4464 samples, and pool and mix every 48 plasma samples into 1 sample. The time to complete the sample addition is 300 minutes;

Step 2: Reagent addition and nucleic acid extraction time is 96 minutes;

Step 3: PCR reagent preparation time is 6 minutes;

Step 4: The PCR reaction system construction time is 13 minutes;

Step 5: PCR sealing and PCR detection time is 80 minutes.

The total testing time for the above process is 495 minutes, or 8.25 hours.

The nucleic acid extraction module of the present application uses a magnetic rod and a cannula to adsorb, separate and transfer magnetic beads, avoiding cross contamination caused by repeated blowing and aspiration mixing with a sample needle or open shaking and mixing with a shaking module. Examples are as follows:

Anti-cross contamination performance verification:

The serum plate consisting of negative and positive samples was used for verification. The serum plate (purchased from Beijing Kangchestein Biotechnology Co., Ltd.) contained: 24 mixed positive samples of HBV and HCV (PC): HBV concentration was 6.6×10 ⁴ IU/mL, HCV concentration was 3.3×10 ⁴ IU/mL, specification: 2mL/tube. 72 negative samples (NC): negative human plasma. The instrument was cross-sampled, and each positive sample was surrounded by negative samples. The sample detection extraction and amplification plate were arranged as shown in Table 1 below.

Table 1

The positive and negative coincidence rates of the test results were statistically analyzed to verify the anti-cross-contamination performance of the present application in detecting strongly positive DNA and RNA samples.

The statistical distribution of the test results is shown in Table 2 below.

Table 2

Note: - represents a negative result, + represents a positive result.

All positive wells tested were positive, with a compliance rate of 24/24; all negative wells tested were negative, with a compliance rate of 72/72, and there was no detection contamination in negative samples, indicating that the application has excellent anti-cross-contamination performance for the detection of strong positive samples of DNA and RNA.

(2) Example of functional modules arranged in two layers

In the embodiment shown in FIG8 , the base 20 is provided with an upper layer and a lower layer, and each functional module is provided in layers. In this embodiment, the sealing module 10 and the PCR detection module 11 are provided in the lower layer of the base, so that the space under the table can be utilized. Reduce the left and right length of the instrument by 30-90cm, reducing the floor space required.

The mechanical gripper can transfer the experimental items through the transfer channel 36 in the table. A mechanical gripper can be shared, or corresponding mechanical grippers can be set up on the upper and lower layers to transfer the experimental items.

The nucleic acid extraction module of the present application uses a magnetic rod sleeve to mix and transfer magnetic beads to extract nucleic acids. The waste liquid is retained in the plate wells. Compared with relying on a pipette to transfer the extracted waste liquid, the influence of residual waste liquid on the purity of nucleic acid extraction is avoided. The instrument of the present application is used in combination with the hepatitis B virus/hepatitis C virus/human immunodeficiency virus (1+2) type nucleic acid detection reagent (purchased from Beijing Wantai Bio-Pharmaceutical Co., Ltd.) to detect the national reference materials of the China Food and Drug Inspection Institute. The HBV DNA detection sensitivity reaches 3IU/mL, the HCV RNA detection sensitivity reaches 10IU/mL, the HIV-1RNA detection sensitivity reaches 40IU/mL, and the HIV-2RNA detection sensitivity reaches 10IU/mL. They are all higher than similar detection systems currently on the market. Examples are verified as follows:

Detection sensitivity performance verification:

The national reference materials of China Food and Drug Inspection Institute were diluted with plasma samples to the following concentrations: HBV DNA 3 IU/mL, HCV RNA 10 IU/mL, HIV-1 RNA 40 IU/mL, HIV-2 RNA 10 IU/mL. The packaging specification is 2 mL/vial. The specific background of the samples is shown in Table 3 below.

Table 3

The instrument of the present application was used with the hepatitis B virus/hepatitis C virus/human immunodeficiency virus (1+2) type nucleic acid detection reagent for detection. 20 samples of each concentration were tested. The detection rate of each concentration sample of each item was counted, and the repeated detection rate of the four items was 100%. The results are shown in Table 4 below.

Table 4

The high-throughput fully automatic nucleic acid detection instrument in this application is divided into a nucleic acid extraction area and a PCR detection area. The PCR detection area is separated independently, and the PCR detection area is isolated by an independent chassis. The PCR detection area is negative pressure, and the PCR detection plate is transmitted through an openable and closable isolation door 9 to prevent the leakage of amplification products from causing pollution in other areas. The air filter cover 16 and the PCR detection area are designed with negative pressure to prevent external dust from interfering with the detection. An exhaust fan 19 and an air filter are provided. The air pressure gradient of the nucleic acid extraction area and the PCR detection area is formed by the exhaust speed and direction of the fan to form a pressure gradient with decreasing air pressure, which meets the reasonable pressure difference setting of nucleic acid detection and prevents pollution during the detection process. An ultraviolet lamp is installed in the air filter cover 16. After the detection is completed, the ultraviolet lamp irradiates each detection area to eliminate nucleic acid contamination in the detection. Solve the defects of the prior art that each functional area has no effective division, air flow has no effective control, and is easy to cause cross contamination of detection.

The waste collection module 141 of the present application is arranged outside the detection area and is only used for discarding pipette tips. The inner layer is a plastic garbage bag and the outer layer is protected by a metal shell to prevent waste from polluting other areas.

In the above embodiments, the description of each embodiment has its own emphasis. For parts that are not described in detail in a certain embodiment, please refer to the detailed description of other embodiments above, and will not be repeated here.

In specific implementation, the above units or structures can be implemented as independent entities, or can be arbitrarily combined to be implemented as the same or several entities. The specific implementation of the above units or structures can refer to the previous method embodiments, which will not be repeated here.

The specific implementation of the above operations can be found in the previous embodiments, which will not be described in detail here.

The above is a detailed introduction to a high-throughput fully automatic nucleic acid detection instrument and method provided in the embodiments of the present application. Specific examples are used in this article to illustrate the principles and implementation methods of the present application. The description of the above embodiments is only used to help understand the method of the present application and its core idea; at the same time, for technical personnel in this field, according to the ideas of the present application, there will be changes in the specific implementation methods and application scopes. In summary, the content of this specification should not be understood as a limitation on the present application.

Claims (11)

A high-throughput fully automatic nucleic acid detection instrument, the high-throughput fully automatic nucleic acid detection instrument comprising: Base (20); A transfer module (4), the transfer module (4) being arranged on the base (20), the transfer module (4) being used to carry a nucleic acid extraction plate, a nucleic acid release plate and a PCR detection plate, wherein the nucleic acid extraction plate, the nucleic acid release plate and the PCR detection plate are all provided with a plurality of plate holes (34), the plate holes (34) of the nucleic acid extraction plate and the nucleic acid release plate being used to receive extraction reagents, and the plate holes (34) of the PCR detection plate being used to receive PCR reaction liquid; A mechanical gripper, wherein the mechanical gripper is used to transport the nucleic acid extraction plate, the nucleic acid release plate and the PCR detection plate; A pipetting module (3), the pipetting module (3) being used to add the test sample into a plurality of plate wells (34) of the nucleic acid extraction plate on the transfer module (4); A nucleic acid extraction module (7), the nucleic acid extraction module (7) comprising a plurality of magnetic head assemblies (30), the magnetic head assemblies (30) comprising a driving device, a sleeve (31), a magnetic rod (35) and magnetic beads (33), the driving device being used to drive the magnetic rod (35) to extend into or out of the sleeve (31), the magnetic rod (35) being used to adsorb or release the magnetic beads (33), the magnetic beads (33) being used to adsorb nucleic acids, when the magnetic rod (35) is extended into the sleeve (31), the magnetic beads (33) are adsorbed onto the outer wall of the sleeve (31), and the magnetic beads (33) follow the sleeve (31) to transfer between plate holes (34); when the magnetic rod (35) is pulled out of the sleeve (31), the magnetic beads (33) adsorbed with nucleic acids are released into the plate holes (34), thereby transferring the nucleic acids; A PCR detection module (11), wherein the PCR detection module (11) is arranged on the base (20), and the PCR detection module (11) is used to perform PCR detection on a PCR detection plate added with nucleic acid to obtain a detection result. According to the high-throughput fully automatic nucleic acid detection instrument according to claim 1, the driving device drives the sleeve (31) to rotate and the solution in the stirring plate hole (34), and the outer wall of the sleeve (31) is provided with corrugations (32). According to the high-throughput fully automatic nucleic acid detection instrument according to claim 1 or 2, the base (20) includes a nucleic acid extraction area, a PCR detection area and an isolation door (9), the isolation door (9) is arranged between the nucleic acid extraction area and the PCR detection area, the pipetting module (3) and the nucleic acid extraction module (7) are located in the nucleic acid extraction area, and the PCR detection module (11) is located in the PCR detection area. According to the high-throughput fully automatic nucleic acid detection instrument according to claim 3, a sealing module (10) is provided on the base (20), and the sealing module (10) is arranged in the PCR detection area, and the sealing module (10) is used to seal the PCR detection plate added with nucleic acid transferred by the mechanical gripper; the PCR detection module (11) is used to detect the PCR detection plate after sealing. According to the high-throughput fully automatic nucleic acid detection instrument according to claim 3, a transmission module (8) is provided between the PCR detection module (11) and the nucleic acid extraction module (7), and the transmission module (8) passes through the isolation door (9), and the isolation door (9) cuts off the transmission module (8) when it is closed, and when the isolation door (9) is opened, the transmission module (8) moves the PCR detection plate added with nucleic acid from the nucleic acid extraction module (7) to the PCR detection module (11). A high-throughput fully-automatic nucleic acid detection instrument according to any one of claims 1-5, wherein a protective rear plate (18) and a protective side plate (17) are vertically arranged on the edge of the base (20), and the protective rear plate (18) and the protective side plate (17) surround the various devices on the base (20). According to claim 6, the high-throughput fully automatic nucleic acid detection instrument, wherein the mechanical gripper and the pipetting module (3) are arranged on the protective rear plate (18), and the base (20) is provided with a barcode scanning module (1), a sample module (2), a consumables carrying position (6) and a reagent module (5), the sample module (2) is located below the pipetting module (3), the sample module (2) is used to carry the detection sample, and the barcode scanning module (1) is used to perform barcode scanning on the detection sample, reagents and consumables in the sample module (2), the transfer module (4), the reagent module (5) and the consumables carrying position (6). According to the high-throughput fully automatic nucleic acid detection instrument according to claim 7, a waste conveying channel (14) is provided on the base (20), and the waste conveying channel (14) is located between the transfer module (4) and the sample module (2), one end of the waste conveying channel (14) extends out of the base (20), and a waste collection module (141) is provided at one end of the waste conveying channel (14), and the waste collection module (141) is located outside the base (20), and a spare consumable position (15) is provided on the protective rear plate (18). According to the high-throughput fully automatic nucleic acid detection instrument according to claim 6 or 7, an air filter cover (16) is provided on the top of the protective rear plate (18) and the protective side plate (17), and an exhaust fan (19) and an air filter are installed on the air filter cover (16), and an ultraviolet lamp is provided in the air filter cover (16), and the ultraviolet lamp is used for disinfection. The high-throughput fully automatic nucleic acid detection instrument according to claim 1, wherein the base (20) is configured as a layer or includes an upper layer and a lower layer, and a transfer channel (36) is provided between the upper layer and the lower layer, and the transfer channel (36) is used to transfer items; The base (20) shares one mechanical gripper, or corresponding mechanical grippers are respectively provided in different areas of the base (20). A high-throughput fully-automatic nucleic acid detection method, characterized in that it is applied to the high-throughput fully-automatic nucleic acid detection instrument according to any one of claims 1 to 10, and the high-throughput fully-automatic nucleic acid detection method comprises: The pipetting module (3) adds the test sample to the multiple plates on the nucleic acid extraction plate on the transfer module (4). Inside the hole (34); The mechanical gripper grabs the nucleic acid extraction plate and places it into the nucleic acid extraction module (7); The plurality of sleeves (31) on the nucleic acid extraction module (7) extend into the plurality of plate holes (34) on the nucleic acid extraction plate for stirring; One or more magnetic rods (35) of the nucleic acid extraction module (7) extend into the sleeve (31), so that the magnetic beads (33) adsorbed with nucleic acid are adsorbed on the outer wall of the sleeve (31); The plurality of magnetic head assemblies (30) of the nucleic acid extraction module (7) are completely separated from the nucleic acid extraction plate; The plurality of magnetic head assemblies (30) of the nucleic acid extraction module (7) are integrally extended into the nucleic acid release plate, and the magnetic rod (35) is pulled out of the sleeve (31) to release the magnetic beads (33) to enter the plurality of plate holes (34) of the nucleic acid release plate; The plurality of sleeves (31) on the nucleic acid extraction module (7) are rotated and stirred in the plurality of plate holes (34) on the nucleic acid release plate, so that the nucleic acid on the magnetic beads (33) is eluted onto the nucleic acid release plate; One or more magnetic rods (35) of the nucleic acid extraction module (7) extend into the sleeve (31), so that the magnetic beads (33) are adsorbed on the outer wall of the sleeve (31); The mechanical gripper moves the nucleic acid release plate carrying the nucleic acid to the transfer module (4); The pipetting module (3) adds the solution in the nucleic acid release plate and the PCR reaction solution to a plurality of plate wells (34) on the PCR detection plate on the transfer module (4) to obtain the PCR detection plate to which nucleic acid is added; The mechanical gripper transports the PCR detection plate with added nucleic acid to the PCR detection module (11) for detection to obtain the detection result.