Nucleic acid extraction and detection all-in-one machine with consumable self-recognition function

Abstract

According to the nucleic acid extraction and detection all-in-one machine with the consumable self-recognition function, through the arrangement of the tip head sensing assembly, the reagent bottle sensing assembly and the eight-connecting-pipe sensing assembly, before formal operation, consumable items in a nucleic acid extraction and amplification kit are subjected to self-recognition detection in sequence; after the completeness and correct placement of consumables of a tip head, a reagent bottle and an eight-connection tube are ensured, nucleic acid extraction and amplification detection is carried out, so that a series of problems caused by incomplete consumables or wrong placement are effectively avoided, and a basic consumable self-identification method and a consumable self-identification method with an automatic comparison and correction function are provided; the method is suitable for consumable self-recognition requirements in different occasions, provides basic guarantee for accurate detection of nucleic acid, is automatically completed in the whole process, and remarkably improves the automation level.

Description

TECHNICAL FIELD

[0001] The present application belongs to the field of biological detection, and relates to a nucleic acid detection device, in particular to a nucleic acid extraction and detection all-in-one machine with self-identification function of consumables. BACKGROUND

[0002] Nucleic acid extraction and amplification detection is an important experimental method in molecular biology research. Early experiments were all completed manually, which was time-consuming and the experimental results were easily affected by individual operation differences of the experimenters. With the development of industrial automation, in the field of experimental science, traditional manual experiments are more and more replaced by automated equipment. The repeatability and consistency of the automated equipment reduce the errors of traditional manual experiments, and improve the reliability and efficiency of the experiments. The nucleic acid extraction and detection all-in-one machine is a high-integration compact instrument, which can automatically and completely extract and amplify nucleic acids, and can highly automatically realize the nucleic acid extraction and amplification detection. The nucleic acid extraction and detection all-in-one machine can shorten the nucleic acid extraction and amplification process, reduce the influence of human factors, enhance the safety and effectiveness of nucleic acid sample preparation, and meet the requirements of miniaturization and portability of the device for rapid detection at the grassroots or on-site, which is the main research direction of developing a new nucleic acid extraction and detection all-in-one machine.

[0003] The nucleic acid extraction and detection all-in-one machine needs to complete the extraction and detection of nucleic acids through a nucleic acid extraction and amplification kit. The nucleic acid extraction and amplification kit is a disposable consumable used in the nucleic acid extraction and detection all-in-one machine. The tip head (a disposable pipette tip), the reagent bottle and the eight-tube are matched consumables pre-placed in the nucleic acid extraction and amplification kit during the production process, and assembled into a complete nucleic acid extraction and amplification kit. However, due to errors in the assembly process or the transportation process, the consumables may be incomplete or the discharge position may be wrong. For example, during the use of the nucleic acid extraction and detection all-in-one machine, sometimes the tip head may not be added to the corresponding position in the nucleic acid extraction and amplification kit, which may cause the machine to stop during the experiment due to insufficient tip heads. The eight-tube may not be placed in the corresponding position in the nucleic acid extraction and amplification kit, or the eight-tube may be incomplete and lack several tubes, which may make the control hole invalid after the experiment, waste the sample and time, cause customer complaints, and even cause the reaction liquid to be added to the inside of the equipment, which is difficult to clean and damages the equipment. In addition, the reagent bottle may be missing or placed in the wrong position, which may affect the subsequent nucleic acid extraction and detection process.

[0004] Therefore, the nucleic acid extraction and detection all-in-one machine must also have a self-identification function of consumables, so as to orderly deal with the above-mentioned sudden problems, avoid invalid detection and waste of time, ensure the accuracy of the detection results, and help protect the nucleic acid extraction and detection all-in-one machine from damage. SUMMARY

[0005] To solve the above problems, the application provides a nucleic acid extraction and detection integrated machine with a consumable self-identification function, which sequentially self-identifies and detects the consumables in a nucleic acid extraction and amplification kit before formal operation, ensures that the consumables of the tip head, reagent bottle and eight-way tube are complete and correctly placed, and then performs nucleic acid extraction and amplification detection, thereby effectively avoiding a series of problems caused by incomplete or incorrectly placed consumables, and providing a basic consumable self-identification method and a consumable self-identification method with automatic comparison and correction function, which is suitable for consumable self-identification needs in different occasions, provides a basic guarantee for accurate nucleic acid detection, and is automatically completed in the whole process, thereby significantly improving the automation level.

[0006] In one aspect, the application provides a nucleic acid extraction and detection integrated machine with a consumable self-identification function, which comprises a nucleic acid extraction and amplification component, a pipetting module and a consumable sensing assembly; the pipetting module is located above the nucleic acid extraction and amplification component and is used to complete the pipetting work of the reagents in the nucleic acid extraction and amplification component; the consumable sensing assembly is used to sense whether the consumables in the nucleic acid extraction and amplification component are in the correct position.

[0007] Further, the consumables in the nucleic acid extraction and amplification component include a tip head, a reagent bottle and an eight-way tube, the consumable sensing assembly includes a tip head sensing assembly, a reagent bottle sensing assembly and an eight-way tube sensing assembly, the tip head sensing assembly is used to sense whether the gun head of the pipetting module is successfully combined with the tip head; the reagent bottle sensing assembly is used to sense whether the reagent bottle is correctly placed; and the eight-way tube sensing assembly is used to sense whether the eight-way tube exists or is complete.

[0008] The tip head of the application is a disposable pipette gun head made of plastic.

[0009] The reagent bottle refers to a bottle used to store the reaction liquid required for nucleic acid extraction and amplification, and the reagents mainly include lysis solution, magnetic bead suspension, washing solution, elution solution, amplification reaction initiator, paraffin oil, paraffin and the like, which are respectively packaged with reagent bottles, sealed with aluminum foil, and numbered on the aluminum foil of different reagent bottles, and placed in a specific position in the nucleic acid extraction and amplification component. The paraffin oil therein is used to seal the reaction tube during nucleic acid amplification reaction, and the paraffin is used to completely seal the liquid in the reaction tube after amplification.

[0010] The eight-way tube is a set of eight plastic reagent tubes connected side by side, each reagent tube contains fluorescently labeled primers for detecting different target nucleic acids, and the tube opening is sealed with aluminum foil.

[0011] The whole process of nucleic acid extraction and amplification reaction needs to go through sample lysis, magnetic bead extraction, washing, elution, amplification, fluorescence detection and other steps, so it will involve multiple taking, changing gun head, puncturing reagent bottle, moving different reagents to different areas for reaction and other complex operations, each step must be accurate, in order to realize the accurate detection of nucleic acid. In this process, the most likely error affecting detection is the missing of consumables, such as the missing of tip head, the wrong placement or missing of reagent bottle, and the missing of eight connected tube, etc. Because the consumables may be missed, lost or wrongly placed during production or transportation, directly used for nucleic acid extraction and detection integrated machine detection, it is very easy to cause a series of problems such as mid-process shutdown, invalid detection, waste of samples, waste of time, reaction liquid entering the internal equipment and being difficult to clean or damaging the equipment.

[0012] In order to completely solve this problem, it is necessary to equip the nucleic acid extraction and detection integrated machine with a consumable self-recognition function to automatically detect whether the consumables have the above problems. The present application sets up respective automatic recognition components for tip head, reagent bottle and eight connected tube: tip head sensing component, reagent bottle sensing component and eight connected tube sensing component, which complete the automatic recognition detection of consumables in all directions. Before starting the nucleic acid detection process, the consumable self-recognition detection is completed according to the setting, ensuring the completeness and correct placement of the consumables, thereby truly providing safety guarantee for subsequent nucleic acid detection.

[0013] Further, the reagent bottle sensing component comprises a capacitance detection device and a gun head; the capacitance detection device is used to detect the capacitance change when the gun head contacts the reagent bottle, so as to judge whether the reagent bottle exists; the gun head is made of metal material, and the reagent bottle is sealed with aluminum foil.

[0014] Since the gun head is made of metal material, the capacitance of the gun head will change when it approaches the aluminum foil, so by detecting the capacitance change of the gun head through the capacitance detection device, it can be directly judged whether the reagent bottle exists, and the next pipetting operation is determined.

[0015] Further, the reagent bottle sensing component further comprises a radio frequency circuit board and a camera component; the radio frequency circuit board is used to detect the RFID information of the nucleic acid extraction and amplification component, to determine the placement order of the reagent bottle in the nucleic acid extraction and amplification component; the camera component is used to shoot the placement order of the reagent bottle, and compare it with the RFID information obtained by the radio frequency circuit board, to judge whether the reagent bottle is placed correctly.

[0016] Using the capacitance change of the gun head to sense whether the reagent bottle exists belongs to the basic consumable self-recognition function provided by the present application, which can be used to judge whether the reagent bottle is missing, but cannot judge whether there is a placement error.

[0017] The basic self-identification function provided by the application includes three aspects: 1. Capacitance change of the gun head to sense whether the reagent bottle exists; 2. Gun head slide sleeve moves up to sense whether the tip head and the gun head are successfully combined; and 3. Fluorescence scanning eight-connected tube detects whether the eight-connected tube exists and is complete.

[0018] On the basis of the basic self-identification function, the self-identification method with automatic comparison and correction function composed of a radio frequency circuit board and a camera assembly is further provided. When the same type of reagent box is used for a long time, and the placement positions of various reagent bottles and tip heads in the reagent box need to be completely fixed, the basic self-identification function can be selected for consumable self-identification; when different types of reagent boxes are often replaced, or the placement positions of various reagent bottles and tip heads in the reagent box need to be corrected, the consumable self-identification with automatic comparison and correction function can be used to provide more comprehensive and higher safety consumable self-identification function.

[0019] The main machine of the nucleic acid extraction and detection all-in-one machine is provided with a storage module and has a memory function, and can perform corresponding pipetting operations according to the stored reagent bottle placement positions, so as to complete the extraction, amplification and detection of nucleic acids.

[0020] In some modes, the nucleic acid extraction and amplification kits in the nucleic acid extraction and amplification component can be of different types, and the placement modes of the reagent bottles and tip heads in different types of kits can be different. The placement modes of the reagent bottles and tip heads in the kit can be determined by the RFID information obtained by the radio frequency circuit board.

[0021] In some modes, the self-identification method with automatic comparison and correction function is that the upper surface of the nucleic acid extraction and amplification kit is photographed by the camera assembly, a photo is obtained, the placement modes of the reagent bottles and tip heads (including the puncture head) in the photo are compared with the placement modes of the reagent bottles and tip heads stored in the reagent box RFID information, and it is judged whether the reagent bottles and tip heads in the photo are missing or have position placement errors. If there is a missing, the operator is reminded through the display screen to supplement the missing consumables as soon as possible at the corresponding missing position; if there is only a position dislocation without a missing, the stored reagent bottle placement positions can be updated according to the existing placement modes, and the operation program is adjusted, so that the subsequent nucleic acid extraction and amplification detection work can be successfully completed.

[0022] Further, the tip head sensing assembly includes a gun head slide sleeve and a sensor, the gun head slide sleeve is movably sleeved outside the gun head, when the gun head takes the tip head, the tip head pushes the gun head slide sleeve to move upward, the upper end of the gun head slide sleeve contacts the sensor to trigger the sensor signal, and it is considered that the tip head and the gun head are successfully combined.

[0023] In some modes, the nucleic acid extraction and amplification component further comprises a puncture head for puncturing the aluminum foil of the reagent bottle before the tip head pipetting.

[0024] Further, the eight-tube sensing assembly comprises a fluorescence detection device for fluorescence scanning of the position where the eight-tube is located, and determining whether the eight-tube exists or is complete according to the scanning result.

[0025] In some modes, according to the fluorescence scanning spectrum, the peak height and the peak value of each peak are calculated and analyzed, if there are specific eight peaks, it is determined that the eight-tube exists, if the number of peaks is insufficient, it is determined to be incomplete, if the peaks do not exist, otherwise it is determined that the eight-tube does not exist.

[0026] Further, the nucleic acid extraction and amplification component comprises a nucleic acid extraction and amplification kit and a bearing module, the bearing module is used for fixing and placing the nucleic acid extraction and amplification kit, the radio frequency circuit board is located in the bearing module and can directly contact with the NFC sticker of the nucleic acid extraction and amplification kit; the pipetting module comprises a displacement assembly, a tip head assembly and a positioning groove, the displacement assembly is provided with an x arm, a y arm and a z arm, and is used for driving the gun head to move in the x, y and z directions; the tip head assembly comprises a gun head, and is used for driving the tip head to complete the pipetting work of the reagent.

[0027] The bearing module can better fix the nucleic acid extraction and amplification kit, and fix the positions of each reagent bottle, tip head, puncture head and eight-tube in the kit. The NFC sticker provides the placement mode of each hole in the kit to the host of the nucleic acid extraction and detection all-in-one machine, and determines the correct placement mode and the subsequent pipetting operation program.

[0028] The nucleic acid extraction and detection all-in-one machine provided by the application mainly realizes the pipetting and reaction of the sample and various reagents in the kit by the pipetting module combined with the tip head and controlling the tip head to complete the pipetting work, and finally realizes the fluorescence detection. Therefore, the pipetting module needs to drive the tip head to realize the accurate displacement in the x, y and z directions, and accurately position the sample to be tested and other reagents in the kit. By setting the displacement assembly comprising the x arm, the y arm and the z arm and improving the structure thereof, the pipetting module can control the gun head to drive the tip head to accurately displace in the x, y and z directions, and minimize the error, so as to better complete the consumable self-recognition work.

[0029] In some modes, the camera assembly comprises a camera, which is arranged at one end of the y-arm track, facilitating the shooting of the entire upper surface of the nucleic acid extraction and amplification kit.

[0030] Further, the positioning groove is located on the upper surface of the bearing module, for positioning the position of the reagent bottle in the nucleic acid extraction and amplification kit; the capacitance detection device is located on the z-arm, for detecting the capacitance change when the gun head approaches the aluminum foil and the capacitance change when the gun head penetrates into the positioning groove; and the camera assembly is located on the y-arm.

[0031] Since the gun head and the positioning groove are also made of metal, the gun head will change the capacitance when approaching the positioning groove, so that the capacitance change of the gun head can be detected by the capacitance detection device, so as to directly judge whether the gun head contacts the positioning groove, thereby helping the positioning of the gun head. Only when the gun head can be precisely positioned, can the consumable self-identification work be better completed.

[0032] In some modes, the positioning groove is a groove with an upward opening, the wall of the groove is cylindrical, and the bottom is hemispherical; the gun head can be inserted into the inside of the groove, and the gun head is calibrated by contacting the wall and the bottom of the groove, so that the specific position of each hole in the reagent kit can be found when performing the pipetting work.

[0033] In some modes, the number of positioning grooves is two. Two points can determine a straight line, and two positioning grooves can precisely calibrate the coordinate position of each reagent bottle in the reagent kit. Meanwhile, two is the minimum number required to complete precise positioning.

[0034] On the other hand, the present application provides a consumable self-identification method of a nucleic acid extraction and detection all-in-one machine, which comprises the following steps:

[0035] (1) placing the nucleic acid extraction and amplification kit into the bearing module of the nucleic acid extraction and detection all-in-one machine;

[0036] (2) sensing whether the eight-way tube exists and is complete by the eight-way tube sensing assembly;

[0037] (3) the gun head positions the position of the reagent bottle through the positioning groove;

[0038] (4) the gun head approaches the reagent bottle, and the capacitance detection device senses whether the reagent bottle exists;

[0039] (5) the gun head is combined with the tip head, and the tip head sensing assembly is used to sense whether the gun head is successfully combined with the tip head.

[0040] Further, step (1) further comprises taking a photo of the upper surface of the nucleic acid extraction and amplification box by the camera assembly, comparing with the stored information, judging whether the tip head and reagent bottle are placed correctly, if it is found that the reagent bottle is placed in wrong order, adjusting the stored information of the reagent bottle and tool placement in the nucleic acid extraction and amplification box, and operating according to the actual placement order.

[0041] The lysis solution reagent bottle, tip head 36, puncture head 16 and the like in the nucleic acid extraction and amplification kit are placed in specific hole positions, and the nucleic acid extraction and detection integrated machine needs to take the tip head, puncture head or remove the reagent from the reagent bottle according to the specific hole position, so the placement of the hole position must be fixed, and if there is a placement error, it will directly affect the subsequent nucleic acid detection.

[0042] However, in actual operation, it is very likely that the reagent bottle in the kit is placed incorrectly, or the tip head is placed incorrectly. Since the reagent bottle is sealed with aluminum foil, the aluminum foil has a number, for example, the lysis solution reagent bottle is marked with No. 5 and needs to be placed in No. 5 hole position, so the placement of the lysis solution reagent bottle can be judged according to the number. The present application directly takes a photo of the upper surface of the nucleic acid extraction and amplification kit by the shooting assembly, and compares the number on the aluminum foil with the hole position where it is placed to determine whether the reagent bottle is placed correctly; after the tip head is placed in the hole position, it can also be determined whether the corresponding hole position has a tip head according to the photo, so as to determine whether the tip head is placed correctly.

[0043] Further, step (1) further comprises obtaining the RFID information of the nucleic acid extraction and amplification kit by the radio frequency circuit board, and storing the information of the reagent bottle and tool placement in the nucleic acid extraction and amplification kit; the camera assembly takes a photo of the upper surface of the nucleic acid extraction and amplification box, compares with the stored information, judges whether the tip head and reagent bottle are placed correctly, if it is found that the reagent bottle is placed in wrong order, adjusts the stored information of the reagent bottle and tool placement in the nucleic acid extraction and amplification kit, and operates according to the actual placement order.

[0044] Some nucleic acid detection projects may need to add or replace different detection reagents or tools, so the kit is provided with multiple backup holes, and the kit is also equipped with different models to cope with it. When replacing different kits, the RFID information of the nucleic acid extraction and amplification kit can be obtained by the radio frequency circuit board, and the information is used to adjust the placement position of the consumables in the kit and store the updated original program. At this time, after taking a photo of the upper surface by the camera assembly, the position information in the photo needs to be compared with the stored information of the reagent bottle and tool placement in the nucleic acid extraction and amplification kit to determine whether there is a placement error of the reagent bottle or tool.

[0045] Further, the eight-tube induction assembly in step (2) senses whether the eight-tube exists and is complete by using a fluorescence detection device to perform fluorescence scanning on the position where the eight-tube is located, and determines whether the eight-tube exists or is complete according to the scanning result.

[0046] In some modes, according to the fluorescence scanning spectrum, the peak height and the peak value of each peak are calculated and analyzed, if there are specific eight peaks, it is determined that the eight-tube exists, if the number of peaks is insufficient, it is determined to be incomplete, and if the peaks do not exist, it is determined that the eight-tube does not exist.

[0047] The nucleic acid extraction and detection all-in-one machine provided by the application has the following beneficial effects:

[0048] (1) By setting the self-identification function of consumables in the nucleic acid extraction and detection all-in-one machine, a series of problems caused by incomplete or incorrect placement of consumables are effectively avoided, providing a basic guarantee for accurate detection of nucleic acids;

[0049] (2) The reagent bottle, tip head (including the puncture head) and eight-tube in the consumables are respectively provided with an induction assembly for self-identification of the consumables in all directions;

[0050] (3) The basic consumable self-identification method and the consumable self-identification method with automatic comparison and correction function are provided, which are suitable for the self-identification needs of consumables in different occasions;

[0051] (4) A pipetting module capable of driving the tip head to accurately displace in x, y and z directions and accurately positioning the hole positions of each consumable is provided, which can minimize the errors that may occur in the pipetting work, so as to better complete the self-identification of the consumables;

[0052] (5) The whole process is automatically completed, which significantly improves the automation level. BRIEF DESCRIPTION OF DRAWINGS

[0053] Figure 1 It is the overall structure schematic diagram of the nucleic acid extraction and detection all-in-one machine with the consumable self-identification function in embodiment 1;

[0054] Figure 2 It is the structure schematic diagram of the nucleic acid extraction and amplification component in embodiment 1;

[0055] Figure 3 It is the split structure schematic diagram of the nucleic acid extraction and amplification component in embodiment 1;

[0056] Figure 4 It is the split structure schematic diagram of the bearing module in embodiment 1;

[0057] Figure 5 It is the structure schematic diagram of the nucleic acid extraction and amplification reagent kit in embodiment 1;

[0058] Figure 6 The exploded view of the nucleic acid extraction and amplification kit in Example 1;

[0059] Figure 7 The schematic diagram of the piercing head structure in Example 1;

[0060] Figure 8 The exploded view of the nucleic acid extraction and amplification kit in Example 1 (from the underside to the top);

[0061] Figure 9 The schematic diagram of the upper surface structure of the nucleic acid extraction and amplification kit in Example 1;

[0062] Figure 10 The schematic diagram of the structure of the pipetting module in Example 1;

[0063] Figure 11 The schematic diagram of the exploded structure of the pipetting module in Example 1;

[0064] Figure 12 The schematic diagram of the structure of the pipetting module in Example 1 (without positioning slot, front view);

[0065] Figure 13 The schematic diagram of the structure of the pipetting module in Example 1 (without positioning slot, side view);

[0066] Figure 14 The schematic diagram of the structure of the tip head assembly in Example 1;

[0067] Figure 15 The exploded view of the tip head sensing assembly in Example 1;

[0068] Figure 16 The cross-sectional view of the tip head assembly in Example 1;

[0069] Figure 17 The schematic diagram of the positioning slot structure in Example 1;

[0070] Figure 18 The schematic diagram of the numbering on the upper surface of the kit in Example 2;

[0071] Figure 19 The fluorescence scanning spectrum when the eight connecting tubes are absent in Example 3;

[0072] Figure 20 The fluorescence scanning spectrum when the eight connecting tubes are complete in Example 3;

[0073] Figure 21 The schematic diagram of the photograph taken by the camera assembly in Example 4. DETAILED DESCRIPTION

[0074] The preferred embodiments of the present invention are further described in detail below in conjunction with the accompanying drawings. It should be noted that the following embodiments are intended to facilitate understanding of the present invention and do not limit it in any way. All features disclosed in the embodiments of the present invention, or all steps in the methods or processes disclosed, except for mutually exclusive features and / or steps, can be combined in any way.

[0075] Example 1: The nucleic acid extraction and detection integrated machine with consumable self-identification function provided by the present invention

[0076] The nucleic acid extraction and detection integrated machine with consumable self-identification function provided in this embodiment is as follows: Figures 1 to 17 shown.

[0077] like Figure 1 The nucleic acid extraction and detection integrated machine 29 with consumables self-identification function provided in this embodiment includes a nucleic acid extraction and amplification component 30, a pipetting module 31 and a consumables sensing component; the pipetting module 31 is located above the nucleic acid extraction and amplification component 30, and is used to complete the pipetting of reagents in the nucleic acid extraction and amplification component 30; the consumables sensing component is used to sense whether the consumables in the nucleic acid extraction and amplification component 30 are in the correct position.

[0078] like Figures 2 to 9 As shown, the nucleic acid extraction and amplification component 30 includes a nucleic acid extraction and amplification reagent kit 32 (hereinafter referred to as the reagent kit 32) and a carrying module 33. The carrying module 33 is used to fix the reagent kit 32. The nucleic acid extraction and detection integrated machine 29 requires the reagent kit 32 to complete the extraction and detection of nucleic acids. The reagent kit 32 is a disposable consumable used in the nucleic acid extraction and detection integrated machine 29. Various consumables are placed in the reagent kit 32 in a specific order. The carrying module 33 can better fix the position of the reagent kit 32, which is equivalent to fixing the position of the various consumables placed in the reagent kit 32.

[0079] Existing nucleic acid extraction and amplification kits typically have poor integration, making it difficult to complete nucleic acid extraction and detection of complex samples. They also have low throughput and can usually only amplify one nucleic acid at a time. When multiple projects are involved, multiple operations are required, which is very time-consuming. The kit 32 provided by the present invention can simultaneously detect multiple projects, but this also makes the various consumables in the kit 32 complex and the order of placement more particular. It must take into account both operational convenience, making the operation less prone to errors, and maintaining balance during placement. The simple and lightweight structure is suitable for large-scale production and transportation.

[0080] like Figures 5 to 9The kit 32 is provided with a sample area 1, a lysis area 3, a waste liquid area 4, a reagent bottle area 34, a tool area 35, and an eight-tube area 44. The sample area 1 is used to place the sample to be tested. The lysis area 3 is used for lysis processing of the sample to be tested, and collects the nucleic acid to be tested in the sample to be tested. The waste liquid area 4 is used to store waste liquid. The reagent bottle area 34 is used to store the reaction liquid required for nucleic acid extraction and amplification, and is sealed with aluminum foil. Different reagent bottles 45 are numbered on the aluminum foil, including a first reagent bottle area 37 and a second reagent bottle area 38. The first reagent bottle area 37 is arranged at the middle position of the kit 32, close to the waste liquid area 8. The first reagent bottle area 37 is provided with a lysis liquid reagent bottle 5, a magnetic bead reagent bottle 7, a first washing liquid reagent bottle 10, a second washing liquid reagent bottle 8, a paraffin oil reagent bottle 11, an amplification reaction initiator reagent bottle 12 (hereinafter referred to as initiator reagent bottle 12), and a backup reagent bottle 13. The second reagent bottle area 38 is provided with an elution liquid reagent bottle 26 and a paraffin reagent bottle 27. Paraffin oil is used to seal the reaction tube during nucleic acid amplification reaction. Paraffin is used to completely seal the liquid in the reaction tube after amplification. The tool area 35 is located between the first reagent bottle area 37 and the second reagent bottle area 38, and is used to store tip heads 36 and puncture heads 16. The tip heads 36 are provided with filter elements, and are arranged at positions of holes 14, 15, 17-24, respectively. The hole 16 is the position of the puncture head 16, and the hole 25 is the backup area. The eight-tube area 44 is used to place eight plastic reagent tubes connected in parallel as a group. Each reagent tube is provided with fluorescently labeled primers for detecting different target nucleic acids. The tube opening is sealed with aluminum foil 46, and is used for amplification and detection of the nucleic acid to be tested. It can be seen that the kit 32 is provided with various consumables such as reagent bottles 45, tip heads 36, puncture heads 16, and eight-tube tubes 28. The types and quantities of consumables are large, and the positional relationship is complex.

[0081] The entire process of nucleic acid extraction and amplification reaction needs to go through multiple steps such as sample lysis, magnetic bead extraction, washing, elution, amplification, and fluorescence detection. Therefore, based on the kit 32, multiple operations such as taking and replacing tip heads 36, puncture heads 16, puncture reagent bottles 45, and moving different reagents to different areas for reaction are required. Each step must be accurate to achieve accurate detection of nucleic acid. In this process, the most likely error affecting detection is the missing of consumables, such as the missing of tip heads 36, the incorrect placement or missing of reagent bottles, and the missing of eight-tube tubes. If the kit 32 is directly used for nucleic acid extraction and detection, it is very easy to cause a series of problems such as machine stop, invalid detection, waste of samples, waste of time, reaction liquid entering the equipment, and difficulty in cleaning or damaging the equipment.

[0082] In order to solve this problem thoroughly, the consumable sensing assembly is equipped on the nucleic acid extraction and detection all-in-one machine 29, so as to have the consumable self-recognition function, and automatically detect whether the consumable has the above problem. The tip head 36, the reagent bottle 45 and the eight-way tube 28 are respectively provided with the respective consumable sensing assembly, which are the tip head sensing assembly 39, the reagent bottle sensing assembly 40 and the eight-way tube sensing assembly 41. The tip head sensing assembly 39 is used to sense whether the gun head 43 of the pipetting module is successfully combined with the tip head 36; the reagent bottle sensing assembly 40 is used to sense whether the reagent bottle 45 is placed correctly; the eight-way tube sensing assembly 41 is used to sense whether the eight-way tube 28 exists or is complete; under the cooperation of the tip head sensing assembly 39, the reagent bottle sensing assembly 40 and the eight-way tube sensing assembly 41, the automatic recognition and detection of the consumable can be completed in all directions. Before starting the nucleic acid detection process, the consumable self-recognition detection is completed according to the setting, so as to ensure the completeness and correct placement of the consumable, thereby truly providing safety guarantee for the subsequent nucleic acid detection.

[0083] As Figure 13 , the reagent bottle sensing assembly 40 includes a capacitive detection device 42 and a gun head 43; the capacitive detection device 42 is used to detect the capacitance change when the gun head 43 contacts the reagent bottle 45, so as to judge whether the reagent bottle 45 exists; the gun head 43 is made of metal material, and the reagent bottle 45 is sealed by an aluminum foil 46. Since the gun head 43 is made of metal material, the gun head 43 will have a capacitance change when it approaches the aluminum foil 46, so that the existence of the reagent bottle 45 can be directly judged by detecting the capacitance change of the gun head through the capacitive detection device 42, and the next pipetting operation is performed after the reagent bottle 45 is determined to exist. The capacitive change of the gun head 43 is used to sense whether the reagent bottle 45 exists, which belongs to the basic consumable self-recognition function, and can be used to judge whether the reagent bottle is missing or not, but cannot judge whether there is a placement error.

[0084] Preferably, the reagent bottle sensing assembly 40 further comprises a radio frequency circuit board 47 and a camera assembly 83; the radio frequency circuit board 47 is located on the side wall of the bearing module 33 and can be in direct contact with the NFC sticker 321 of the reagent box 32, for detecting the RFID information of the reagent box 32 and determining the placement order of the consumables in the reagent box 32; the camera assembly 83 comprises a camera 49 and is arranged at one end of the track of the y-arm 50 of the pipetting module 31, facilitating the shooting of the entire upper surface of the reagent box 32, for shooting a photo of the placement order of the consumables and comparing with the RFID information obtained by the radio frequency circuit board 47 to determine whether the consumables in the reagent box 32 are placed correctly. The radio frequency circuit board 47 and the camera assembly 83 can provide self-identification with automatic comparison and correction function. The main machine of the nucleic acid extraction and detection all-in-one machine 29 is provided with a storage module and has a memory function, which can perform corresponding pipetting operations according to the stored placement positions of the reagent bottles 45, thereby completing the extraction, amplification and detection of nucleic acids. There can be different models of nucleic acid extraction and amplification reagent boxes 32, and the placement modes of the reagent bottles 45 and the tip heads 36 in different models of reagent boxes 32 can be different. The RFID information obtained by the radio frequency circuit board 47 can determine the placement mode of the reagent bottles 45 and the tip heads 36 in the reagent box 32. When the same model of reagent box 32 is used for a long time and the placement positions of various reagent bottles 45 and tip heads 36 in the reagent box 32 need to be completely fixed, the basic self-identification function can be selected for consumable self-identification; when different models of reagent boxes 32 need to be replaced or the placement positions of various reagent bottles 45 and tip heads 36 in the reagent box 32 need to be corrected, the consumable self-identification with automatic comparison and correction function can be used to provide more comprehensive and higher safety consumable self-identification function.

[0085] As Figures 10 to 16 The tip head sensing assembly 39 is located on the pipetting module 31 and comprises a gun head sliding sleeve 48 and a sensor 49. The gun head sliding sleeve 48 is movably sleeved outside the gun head 43. When the gun head 43 takes the tip head 36, the tip head 36 pushes the gun head sliding sleeve 48 to move upward, and the trigger light coupling 481 at the upper end of the gun head sliding sleeve 48 contacts the sensor 49, triggering the sensor signal and considering that the tip head 36 is successfully combined with the gun head 43. The consumables in the reagent box 32 further comprise a piercing head 16 for piercing the aluminum foil 46 of the reagent bottle 45, and then the pipetting work of the tip head 36 can be performed. The tail end of the piercing head 16 has the same specification as the tip head 36 and also needs to be combined with the gun head 43 to complete the work of piercing the aluminum foil 46 under the control of the gun head 43, so the tip head sensing assembly 39 can also be used to sense whether the combination is successful.

[0086] As Figure 10, the displacement assembly 51 drives the tip head assembly 52 to displace, thereby driving the tip head 36 to displace through the gun head 43; when the tip head 36 is separated from the gun head 43, the tip head assembly 52 drives the ejecting fork rod 72 to displace, and under the limitation of the ejecting fork rod 72, the tip head 36 stops upward movement, thereby separating the tip head 36 from the gun head 43 which continues to move upward. The ejecting fork rod 72 is controlled by the electromagnet 73 to displace or retract; when the electromagnet 73 is electrified from one direction, magnetism is generated, causing the ejecting fork rod 72 to be pushed out due to repulsion of the same polarity; when the electromagnet 73 is electrified in the opposite direction, opposite magnetism is generated, causing the ejecting fork rod 73 to be retracted due to attraction of the different polarity.

[0087] As Figures 14 to 16 , the upper end of the gun head 43 is provided with a limiting device 79 of the gun head sliding sleeve 48, and the limiting device 79 includes a spring 80 and a baffle 81; the upper end of the spring 80 is fixed to the outside of the upper end of the gun head 43, and is used to block the upward movement of the gun head sliding sleeve 48; the baffle 81 can abut against the edge 82 of the upper end of the gun head sliding sleeve 48 to prevent the gun head sliding sleeve 48 from moving downward to separate from the gun head 43. The limiting device 79 can limit the highest and lowest positions of the gun head sliding sleeve 48, so that the gun head sliding sleeve 48 is always outside the gun head 43 and wraps the gun head 43, and cannot fall off; meanwhile, the design of the spring 80 can provide a certain range of upward movement space for the gun head sliding sleeve 48. When the tip head 36 is combined with the gun head 43, the gun head sliding sleeve 48 can be pushed to move upward, and the spring 80 is compressed; when the tip head 36 is separated from the gun head 43, the gun head sliding sleeve 48 moves downward due to gravity and is fixed by the baffle 81 to be in the lowest position, and the spring 80 returns to normal.

[0088] As Figures 10 to 13 , the displacement assembly 51 of the pipetting module 31 is provided with an x arm 54, a y arm 50 and a z arm 55, which are respectively used to drive the gun head 43 to move in the x, y and z directions; the tip head assembly 52 includes the gun head 43, which is used to drive the tip head 36 to complete the pipetting work of the reagent. The nucleic acid extraction and detection all-in-one machine 29 mainly realizes the pipetting and reaction of the samples and various reagents in the reagent box 32 through the combination of the pipetting module 31 and the tip head 36, and controls the tip head 36 to complete the pipetting work, and finally realizes the fluorescence detection. Therefore, the pipetting module 31 needs to be able to drive the tip head 36 to realize the precise displacement in the x, y and z directions, and to be able to accurately position the samples and other reagents to be tested in the reagent box 32. By setting the displacement assembly including the x arm, the y arm and the z arm, and improving the structure, the pipetting module can control the precision of the displacement of the gun head driving the tip head in the x, y and z directions, and can minimize the possible errors, thereby better completing the consumable self-recognition work.

[0089] As Figures 11 to 13 , in order to realize the precise displacement of the tip head 36 in the three-dimensional space, the x-arm 54 is located at the top; the y-arm 50 is located below the x-arm 54, and the x-arm 54 can drive the y-arm 50 to displace in the x direction; the z-arm 55 is located below the y-arm 50, and the y-arm 50 can drive the z-arm 55 to displace in the y direction. The x-arm 54 and the y-arm 50 are horizontally placed, the y-arm 50 is located below the x-arm 54 and horizontally crosses the x-arm 54 at a 90-degree angle, and the z-arm 55 is vertically placed below the y-arm 50. As Figure 9 and 10 , the x-arm 54, the y-arm 50 and the z-arm 55 are respectively provided with a stepping motor, a guide rail, a synchronous belt and a zero sensor. Through the setting of the stepping motor, the guide rail and the synchronous belt, the precision of displacement in the x, y and z directions can be controlled, and the initial position is controlled through the zero sensor to minimize errors. Among them, the x-arm 54 is provided with a first stepping motor 56, a first guide rail 57 and a first synchronous belt 58; the y-arm 50 is provided with a second stepping motor 59, a second guide rail 60 and a second synchronous belt 61; and the z-arm 55 is provided with a third stepping motor 62, a third guide rail 63 and a third synchronous belt 64. The zero sensor includes a first zero sensor 65 (x-direction zero sensor), a second zero sensor 66 (y-direction zero sensor) and a third zero sensor 67 (z-direction zero sensor), which are used to sense the starting positions in the x, y and z directions respectively. Each zero sensor is composed of a baffle and a sensor; among them, the first zero sensor 65 in the x direction, the first sensor 66 is located on the x-arm 54, and the first baffle 67 is located on the y-arm 50; the second zero sensor 66 in the y direction, the second sensor 68 is located on the z-arm 55, and the second baffle 69 is located on the y-arm 50; the third zero sensor 67 in the z direction, the third sensor 70 is located on the z-arm 55, and the third baffle 71 is also located on the z-arm 55. The zero sensors in the x, y and z directions are respectively cross-set, which can be more conducive to simultaneously controlling the positional relationship among the x-arm 54, the y-arm 50 and the z-arm 55, and the structure design is also more compact. The tip head assembly 52 is located on the z-arm 55 and is driven by the z-arm 55 to displace up and down in the z direction; the y-arm 50 drives the z-arm 55 to displace in the y direction; and the x-arm 54 drives the y-arm 50 to displace in the x direction.

[0090] The capacitive detection device 42 is located on the z arm 55, which is used to detect the capacitive change when the gun head 43 contacts the reagent bottle 45, so as to determine whether the reagent bottle 45 exists, and is also used to detect the capacitive change when the gun head 43 contacts the positioning groove 53; the gun head 43 and the positioning groove 53 are made of metal, and the gun head 43 will change the capacitance when it is close to the positioning groove 53. Therefore, by detecting the capacitive change of the gun head 43 through the capacitive detection device 42, it can be directly determined whether the gun head 43 contacts the positioning groove 53, so as to help the gun head 43 accurately position the hole position of each consumable in the reagent box 32. Through the positioning of the positioning groove 53, it is also helpful to better complete the self-identification of the consumables.

[0091] As Figure 10 , the positioning groove 53 is located on the upper surface of the bearing module 33, which is an open upward groove. The wall 74 of the groove body is cylindrical, and the bottom 75 is hemispherical, which is fixed on the upper surface of the bearing module 33 by a screw 76. The gun head 43 can be inserted into the inside of the groove body and be calibrated by contacting the wall 74 and the bottom 75 of the groove body, so that the position of the reagent can be found when the pipetting work is performed. The top end of the gun head 43 is designed in a spherical shape, and the size of the positioning groove 53 can match or be slightly larger than the size of the gun head 43, so that the gun head 43 can be inserted into the inside of the positioning groove 53. Before the gun head 43 is combined with the tip head 36 to perform the pipetting work, it first detects the position of the positioning groove 53 for accurate positioning, and then performs the combination with the tip head 36 and completes the pipetting work. The positioning groove 53 is equivalent to calibrating the position of the sample reagent. By determining the position of the positioning groove 53, the gun head 43 can calculate the specific hole position of various reagents in the reagent box 32, help the gun head 43 accurately position, and accurately complete the complex pipetting work. The number of the positioning groove 53 is two, because two points can determine a straight line, and two positioning grooves 53 can accurately calibrate the coordinate position of the reagent box 32, so as to determine the specific hole position of various reagent bottles in the reagent box 32. At the same time, two positioning grooves 53 are also the minimum number required to complete accurate positioning.

[0092] The plunger pump 77 is also provided on the pipetting module 31 Figures 11 to 12 , which is located below the y arm 50 and is connected with the gun head 43 through a pipeline, and is used to control the adsorption or discharge of the reagent by the tip head 36, and also can control the amount of adsorbed reagent.

[0093] As Figure 1 , the eight-tube sensing assembly 41 includes a fluorescence detection device 78, which is used to perform fluorescence scanning on the position where the eight-tube 28 is located. According to the fluorescence scanning spectrum, the values of the peak height and the peak valley of each peak are calculated and analyzed, and the scanning result is used to determine whether the eight-tube 28 exists or is complete: if there are specific 8 peaks, it is determined that the eight-tube exists, if the number of peaks is insufficient, it is determined to be incomplete, and if the peak does not exist, it is determined that the eight-tube does not exist.

[0094] In summary, the nucleic acid extraction and detection all-in-one machine 29 provided in the embodiment has the following three aspects of self-identification functions of basic consumables: 1. The capacitance change of the gun head 43 is used to sense whether the reagent bottle 45 is present; 2. The upward movement of the gun head sliding sleeve 48 is used to sense whether the tip head 36 and the gun head 43 are successfully combined; and 3. The fluorescence scanning eight-connection tube 28 is used to detect whether the eight-connection tube is present and complete. At the same time, since the pipetting module 31 can drive the tip head 36 to accurately displace in the x, y and z directions, and can accurately position the hole positions of the consumables through the positioning groove 53, the errors that may occur in the pipetting work are minimized, so that the self-identification work of the consumables can be better completed.

[0095] On the basis of the basic self-identification function, the self-identification method of the consumables with the automatic comparison and correction function can also be realized: the upper surface of the reagent box 32 is photographed by the camera assembly 83, the photograph is obtained, the arrangement mode of the reagent bottle 45 and the tip head 36 (including the puncture head 16) in the photograph is compared with the arrangement mode of the reagent bottle 45 and the tip head 36 stored in the RFID information of the reagent box 32, and it is judged whether the reagent bottle 45 and the tip head 36 in the photograph are missing or have a position arrangement error. If there is a missing, the operator is reminded through the display screen to supplement the missing consumables as soon as possible in the corresponding missing position; if there is only a position misplacement without a missing, the stored arrangement position can be updated according to the existing arrangement mode, and the operation program is adjusted, so that the subsequent nucleic acid extraction and amplification detection work can be smoothly completed.

[0096] Embodiment 2, nucleic acid extraction and detection method of nucleic acid extraction and detection all-in-one machine

[0097] In this embodiment, the nucleic acid extraction and detection all-in-one machine 29 provided in Embodiment 1 is used for nucleic acid extraction and amplification and detection, and the nucleic acid extraction and detection method of the nucleic acid extraction and detection all-in-one machine 29 is used for reference. Figure 18 , and specifically includes the following steps:

[0098] 1. Place the reagent box 32 into the nucleic acid extraction and detection all-in-one machine 29 and fix it, click start detection according to the prompt operation, the instrument performs self-checking and confirmation, and the lysis area 3 starts heating;

[0099] 2. The pipetting gun of the nucleic acid extraction and detection all-in-one machine takes the puncture head 16 from the 16-hole position of the tool area 35, and breaks the film of the reagent bottle in the first reagent bottle area 37;

[0100] 3. The gun head 43 takes the tip head 36 at the hole 14 position, moves to the hole position of the sample area 1, takes the sample, and moves to the lysis area 3 to release the sample;

[0101] 4. Move to the lysis liquid reagent bottle 5 to take the lysis liquid, move to the lysis area 3 to release the lysis liquid;

[0102] 5. Gun head 43 moves to magnetic bead reagent bottle 7 hole position, takes magnetic beads, moves to lysis area 3, releases magnetic beads, gun head 43 moves to designated position, after the end of the step, returns TIP head to hole 14 position, starts magnetic attraction (through the magnetic attraction of magnet 2, the magnetic beads are gathered in small space 84 of lysis area 3), lysis area 3 stops heating;

[0103] 6. Gun head 43 moves to hole 14 position to take TIP head, moves to lysis area 3, and transfers the supernatant to waste liquid area 4;

[0104] 7. Gun head 43 moves to the first washing liquid reagent bottle 10 hole position, absorbs the first washing liquid and transfers it to the lysis area 3, and releases the washing liquid; after the end of the step, the supernatant is transferred to the waste liquid area 4;

[0105] 8. Gun head 43 moves to the second washing liquid reagent bottle 8 hole position, takes the second washing liquid, and transfers it to the lysis area 3, and releases the second washing liquid; after the end of the step, the supernatant is transferred to the waste liquid area 4;

[0106] 9. Return TIP head to hole 14 position, release magnetic attraction;

[0107] 10. Gun head 43 moves to 16 hole position to take puncture head 16, punctures the sealing film of eluent reagent bottle 26 and eight-way tube 28, and returns puncture head 16 after the end of the step; moves to hole 15 position to take TIP head 36, moves to eluent reagent bottle 26 hole position, takes eluent and places it in each tube of eight-way tube 28 (including 281-288, for dissolving freeze-dried powder in each eight-way tube);

[0108] 11. Gun head 43 moves to eluent reagent bottle 26 hole position, takes eluent, moves to lysis area 3, and releases eluent; after the end of the step, gun head 43 moves to eluent reagent bottle 26 hole position to clean tip head;

[0109] 12. Gun head 43 moves to hole 15 position to return tip head, and starts magnetic attraction;

[0110] 13. Gun head 43 moves to hole 17 position to take tip head, moves to lysis area 3, takes purified nucleic acid, moves to starting agent reagent bottle 12 position to release, and after the end of the step, gun head 43 moves to hole 17 position to return tip head;

[0111] 14. Gun head 43 moves to hole 18 position to take tip head, moves to starting agent reagent bottle 12 hole position to take nucleic acid, and then moves to the second tube 282 of eight-way tube 28 to release nucleic acid; after the end of the step, gun head 43 moves to hole 18 position to return tip head;

[0112] 15. Repeat step 14 for tip head in hole 19, hole 20, hole 21, hole 22, hole 23, hole 24 position to transfer nucleic acid into the third tube 283, the fourth tube 284, the fifth tube 285, the sixth tube 286, the seventh tube 287, the eighth tube 288 of the eight-tube 28 respectively (each time the tip head in hole 19, hole 20, hole 21, hole 22, hole 23, hole 24 position is changed to avoid cross contamination); Figure 8

[0113] 16. The gun head 43 moves to the hole 15 position to take the tip head, moves to the paraffin oil reagent bottle 11 hole position, takes the paraffin oil, and returns the tip head after releasing the paraffin oil;

[0114] 17. Start collecting fluorescence in the eight-tube 28;

[0115] 18. The gun head 43 moves to the hole 14 position to take the tip head, moves to the lysis area 3, sucks the residual liquid, moves to the waste area 4, and returns the tip head after the step is completed;

[0116] 19. After 36 times of fluorescence collection (40 times of continuous fluorescence collection are required for each hole, and the paraffin sealing is started at the 36th time), the pipette moves to the puncture head hole position 16 to take the puncture head 16, and the paraffin reagent bottle 27 is punctured at the paraffin reagent bottle 27 hole position; return the tip head after the step is completed;

[0117] 20. The gun head 43 moves to the hole 15 position to take the tip head, moves to the paraffin reagent bottle 27 hole position, takes the paraffin, and moves to the eight-tube 28; return the TIP head after the step is completed;

[0118] 21. Stop heating the paraffin area, and after the fluorescence collection is completed, the nucleic acid extraction, amplification and detection of this time are completed.

[0119] The 21-step procedure is stored in the nucleic acid extraction and detection all-in-one machine 29, and each time the nucleic acid extraction and detection work is completed according to the procedure.

[0120] Example 3, basic consumable self-identification method

[0121] This embodiment uses the nucleic acid extraction and detection all-in-one machine 29 provided in example 1 to perform basic consumable self-identification, and the method is as follows:

[0122] (1) After the reagent box 32 is unpacked, it is placed in the bearing module 33 of the nucleic acid extraction and detection all-in-one machine 28;

[0123] (2) The eight-tube sensing assembly 42 senses whether the eight-tube 28 exists and is complete:

[0124] ​The fluorescence detection device 78 first scans the eight-tube area 44 in the kit 32 and obtains scanning data, and determines whether the eight-tube 28 exists and is complete according to the following method, and if not, timely reminds the user to check and replace the kit 32.

[0125] If the eight-tube 28 does not exist, the fluorescence scanning spectrum is as follows: Figure 19 , and no peak appears after the starting point of the abscissa; if the eight-tube 28 exists and is complete, the fluorescence scanning spectrum is as follows: Figure 20 , and the peak height threshold is set to a. According to the device positioning, the ordinate corresponding to the first peak valley is marked as y1, and the abscissa x1 is found. The maximum value of the ordinate between the first peak valley and the second peak valley is calculated and marked as y2, and the corresponding abscissa x2 is calculated. The peak height h=y2-y1 and the peak width b=x2-x1 are calculated.

[0126] The ordinate of the abscissa x1+b / 3 is marked as y3.

[0127] The ordinate of the abscissa x1+b*2 / 3 is marked as y4.

[0128] The ordinate of the abscissa x1+b*4 / 3 is marked as y5.

[0129] The ordinate of the abscissa x1+b*5 / 3 is marked as y6.

[0130] At the same time, the following conditions are met:

[0131] y2>y4>y3>y1 (only greater than, it is considered that the eight-tube exists, otherwise it is considered that the determination fails);

[0132] y2>y5>y6>y1 (only greater than, it is considered that the eight-tube exists, otherwise it is considered that the determination fails);

[0133] h>a (h is higher than the peak height threshold a, and it is considered that the peak exists);

[0134] In this way, if the first 8 peaks exist, it is determined that the eight-tube 28 exists, otherwise it is determined that the eight-tube 28 does not exist, or lacks some peaks, and it is determined that the eight-tube 28 is not complete.

[0135] (3) The gun head 43 positions the position of the reagent bottle 45 through the positioning slot 53:

[0136] The gun head 43 moves above one of the positioning slots 53 and probes into the positioning slot 53. When the gun head 43 contacts the wall 74 or the bottom 75 of the positioning slot 53, the capacitance detection device detects an increase in capacitance (greater than 1 pf), and the positioning of one positioning slot is completed. Then the gun head is moved above another positioning slot 53 and probes into it. After the positioning of two positioning slots is completed, the exact position of the reagent box 32 is determined, and the specific hole positions of all consumables in the reagent box 32 are determined.

[0137] (4) The gun head 43 approaches the reagent bottle 45, and the capacitance detection device 42 senses whether the reagent bottle 45 is present:

[0138] After positioning, the gun head 43 is moved above the reagent box 32. When the gun head 43 is located directly above the reagent bottle 45 in a certain hole position and contacts the sealing aluminum foil of the reagent bottle in the hole position, the capacitance detection device detects an increase in capacitance (greater than 1 pf), indicating that the reagent bottle in the hole position is present. Conversely, when the gun head 43 is located directly above the reagent bottle 45 in a certain hole position and contacts the sealing aluminum foil 46 of the reagent bottle in the hole position, the capacitance detection device does not detect a change in capacitance, indicating that the reagent bottle in the hole position is not present.

[0139] (5) The gun head 43 is combined with the tip head 36, and the tip head sensing assembly 39 is used to sense whether the gun head 43 is successfully combined with the tip head 36:

[0140] After determining that the reagent bottles 45 are correctly placed, the gun head 43 moves to the tool area 35 of the reagent box 32, finds the correct hole position of the tip head 36, and then the gun head 43 descends and is combined with the tip head 36. At this time, the tip head 36 pushes the gun head slide sleeve 48 to move upward, and the trigger light coupling 481 at the upper end of the gun head slide sleeve 48 contacts the sensor 49, triggering the sensor signal and indicating that the tip head 36 is successfully combined with the gun head 43.

[0141] After the basic consumable self-identification is completed and all requirements are met, the nucleic acid extraction and detection all-in-one machine 29 begins the formal nucleic acid extraction, amplification, and detection process as provided in Embodiment 2.

[0142] Embodiment 4, Consumable self-identification method with automatic comparison and correction function

[0143] This embodiment uses the nucleic acid extraction and detection all-in-one machine 29 provided in Embodiment 1 to perform consumable self-identification with an automatic comparison and correction function.

[0144] As Figure 18As shown, the lysate reagent bottle 5 (No. 5 hole position), the magnetic bead reagent bottle 7 (No. 7 hole position), the first washing liquid reagent bottle 10 (No. 10 hole position), the second washing liquid reagent bottle 8 (No. 8 hole position), the paraffin oil reagent bottle 11 (No. 11 hole position), the initiator reagent bottle 12 (No. 12 hole position), the backup reagent bottle 13 (No. 13 hole position) in the first reagent bottle area 37, the eluent reagent bottle 26 (No. 26 hole position) and the paraffin reagent bottle 27 (No. 27 hole position) in the second reagent bottle area 20, the tip head 36 in the hole 14, 15, 17-24 position, the puncture head 16 (No. 16 hole position), the backup hole 25 (No. 25 hole position) in the tool area 35, and the eight-way tube 28 of the eight-way tube area 44 are placed in the specific hole position. The nucleic acid extraction and detection integrated machine 29 must take the tip head, the puncture head or the reagent from the reagent bottle according to the specific hole position, so the placement of the hole position must be fixed. If the placement is wrong, it will directly affect the nucleic acid detection. Therefore, the nucleic acid extraction and detection integrated machine with the automatic comparison correction function of the consumable self-identification method is provided, which takes a photo of the upper surface of the reagent box by the camera assembly to check the placement error of the reagent bottle and the tool, thereby avoiding a series of problems caused by the placement error.

[0145] I. Automatic comparison correction function of consumable error placement

[0146] Figure 18 The correct placement of the consumables in the reagent box 32 is shown. If the placement is wrong, there may be the following situations:

[0147] 1. The reagent bottle placement sequence is moved back as a whole

[0148] Due to the error in the reagent bottle placement sequence, all the reagent bottles are moved back as a whole, the No. 5 hole position is empty, the lysate reagent bottle is in the No. 6 hole position, the magnetic bead reagent bottle is in the No. 8 hole position, the first washing liquid reagent bottle is in the No. 11 hole position, the second washing liquid reagent bottle is in the No. 9 hole position, the paraffin oil reagent bottle is in the No. 12 hole position, and the initiator reagent bottle is in the No. 13 hole position. At this time, the photo of the upper surface of the reagent box taken by the camera assembly can determine that the above-mentioned reagent bottles are moved back as a whole, and the nucleic acid extraction and detection integrated machine can adjust the original operation program (the operation program in embodiment 2) according to the position of the reagent bottle in the taken photo, and move back the hole position in the original operation program as a whole. Therefore, at this time, it is not necessary to stop to adjust the reagent bottle placement sequence, and the subsequent nucleic acid extraction, amplification and detection work can still be smoothly completed.

[0149] 2. The placement position of a certain reagent bottle is wrong, but there is no missing reagent bottle

[0150] The lysis solution reagent bottle is placed in the No. 6 hole, and other reagent bottles are placed correctly. At this time, the photo of the upper surface of the reagent box taken by the camera assembly can determine that the lysis solution reagent bottle is placed in the No. 6 hole. The nucleic acid extraction and detection all-in-one machine can adjust the original operation program according to the position of the lysis solution reagent bottle in the photographed photo, and adjust the hole position of the lysis solution reagent bottle in the original operation program from No. 5 to No. 6. Therefore, at this time, it is not necessary to stop the machine to adjust the placement order of the lysis solution reagent bottle, and the subsequent nucleic acid extraction, amplification and detection work can still be completed smoothly.

[0151] Similarly, when multiple reagent bottle positions are placed incorrectly, the operation program can also be adjusted to correct the error without stopping the machine, and the nucleic acid extraction, amplification and detection work can still be carried out normally.

[0152] 3. Lack of tip head in a certain hole

[0153] The tip head in the No. 14 hole of the reagent box is missing, but the backup tip head is placed in the backup hole No. 25. At this time, the photo of the upper surface of the reagent box taken by the camera assembly can determine that the tip head in the No. 14 hole is missing. The nucleic acid extraction and detection all-in-one machine can adjust the original operation program according to the photographed photo, and modify the program of taking the tip head from the No. 14 hole in the original operation program to taking the tip head from the No. 25 hole. Therefore, it is not necessary to stop the machine to supplement the tip head, and the subsequent nucleic acid extraction, amplification and detection work can still be completed smoothly.

[0154] 4. Lack of reagent bottle

[0155] The photo of the upper surface of the reagent box taken by the camera assembly shows that a certain reagent bottle is missing. At this time, the nucleic acid extraction and detection all-in-one machine will automatically stop and issue a warning to remind the operator to supplement the missing reagent bottle before the subsequent nucleic acid extraction, amplification and detection work can be carried out.

[0156] II. Reagent box model change adjustment function

[0157] 1. Add new reagent bottle

[0158] When a new reagent box model needs to be replaced, the RFID information of the reagent box is first obtained through the radio frequency circuit board, and the placement position of the consumables in the reagent box is adjusted according to the information and the original program is updated. For example, in the new reagent box, the No. 9 hole is placed with a lysis solution additive. At this time, the nucleic acid extraction and detection all-in-one machine needs to adjust the original program according to the RFID information of the reagent box to update the operation program, so as to carry out the subsequent nucleic acid extraction, amplification and detection work.

[0159] 2. Incorrect placement of reagent bottle

[0160] After replacing the new kit model, the nucleic acid extraction and detection all-in-one machine obtains the RFID information of the kit through the radio frequency circuit board, adjusts the placement position of the consumables in the kit according to the information, and stores the original program. After taking a photo of the upper surface through the camera assembly, the position information in the photo needs to be compared with the placement information of the reagent bottles and tools in the newly stored kit to determine whether there is a placement error of the reagent bottles or tools. If there is a placement error of the reagent bottles, it can also be corrected by adjusting the operation program without stopping the machine, and the nucleic acid extraction, amplification and detection can still be carried out normally.

[0161] Figure 21 The schematic diagram for the camera assembly to take a photo, in actual operation, the reagent bottle numbers in the kit are different from those in the present application, but the basic principles are consistent.

[0162] In addition, even if the nucleic acid extraction and detection all-in-one machine has been automatically compared and corrected, it still needs to perform basic consumable self-identification before formal work, which can provide more protection for subsequent nucleic acid extraction and detection.

[0163] The application of the present application is not limited to this. For example, its application range in environmental protection can be extended. Any person skilled in the art can make various changes and modifications without departing from the spirit and scope of the present application. Therefore, the protection scope of the present application should be limited by the scope defined in the claims.

Claims

1. A nucleic acid extraction and detection integrated machine with consumables self-identification function, characterized in that: It includes a nucleic acid extraction and amplification component, a pipetting module and a consumables sensing component; the pipetting module is located above the nucleic acid extraction and amplification component and is used to complete the pipetting of reagents in the nucleic acid extraction and amplification component; the consumables sensing component is used to sense whether the consumables in the nucleic acid extraction and amplification component are in the correct position.

2. The nucleic acid extraction and detection integrated machine according to claim 1, characterized in that: The consumables in the nucleic acid extraction and amplification component include a tip head, a reagent bottle and an eight-tube connector. The consumables sensing assembly includes a tip head sensing assembly, a reagent bottle sensing assembly and an eight-tube connector sensing assembly. The tip head sensing assembly is used to sense whether the gun head of the pipetting module is successfully combined with the tip head; the reagent bottle sensing assembly is used to sense whether the reagent bottle is placed correctly; and the eight-tube sensing assembly is used to sense whether the eight-tube connector exists or is complete.

3. The nucleic acid extraction and detection integrated machine according to claim 2, characterized in that: The reagent bottle sensing assembly includes a capacitance detection device and a gun head; the capacitance detection device is used to detect the capacitance change when the gun head contacts the reagent bottle, thereby determining whether the reagent bottle exists; the gun head is made of metal, and the reagent bottle is sealed with aluminum foil.

4. The nucleic acid extraction and detection integrated machine according to claim 3, characterized in that: The reagent bottle sensing component also includes a radio frequency circuit board and a camera component; the radio frequency circuit board is used to detect the RFID information of the nucleic acid extraction and amplification component and determine the placement order of the reagent bottles in the nucleic acid extraction and amplification component; the camera component is used to photograph the placement order of the reagent bottles and compare it with the RFID information obtained by the radio frequency circuit board to determine whether the reagent bottles are placed correctly.

5. The nucleic acid extraction and detection integrated machine according to claim 4, characterized in that: The tip head sensing assembly includes a gun head slide and a sensor. The gun head slide can be movably mounted on the outside of the gun head. When the gun head takes the tip head, the tip head pushes the gun head slide to move upward. The upper end of the gun head slide contacts the sensor, triggering the sensor signal, indicating that the tip head and the gun head are successfully combined.

6. The nucleic acid extraction and detection integrated machine according to claim 5, characterized in that: The eight-tube sensing assembly includes a fluorescence detection device, which is used to perform fluorescence scanning on the location of the eight-tube and determine whether the eight-tube exists or is complete based on the scanning result.

7. The nucleic acid extraction and detection integrated machine according to claim 6, characterized in that: The nucleic acid extraction and amplification component includes a nucleic acid extraction and amplification kit and a carrying module. The carrying module is used to fix the nucleic acid extraction and amplification kit. The radio frequency circuit board is located in the carrying module and can directly contact the NFC sticker of the nucleic acid extraction and amplification kit; the pipetting module includes a displacement component, a tip head component and a positioning groove. The displacement component is provided with an x-arm, a y-arm and a z-arm, which are used to drive the displacement movement of the gun head in the x, y and z directions; the tip head component includes a gun head, which is used to drive the tip head to complete the pipetting of the reagent.

8. The nucleic acid extraction and detection integrated machine according to claim 7, characterized in that: The positioning groove is located on the upper surface of the carrying module and is used to locate the position of the reagent bottle in the nucleic acid extraction and amplification kit; the capacitance detection device is located on the z arm and is used to detect the capacitance change when the gun tip approaches the aluminum foil and the capacitance change when the gun tip extends into the positioning groove; the camera assembly is located on the y arm.

9. A method for self-identification of consumables of a nucleic acid extraction and detection integrated machine, characterized in that: The following steps are involved: (1) Place the nucleic acid extraction and amplification kit into the carrier module of the nucleic acid extraction and detection integrated machine; (2) Using the eight-tube sensing component to sense whether the eight-tube exists and is complete; (3) The gun tip locates the position of the reagent bottle through the positioning groove; (4) The gun tip is brought close to the reagent bottle, and the capacitance detection device senses whether the reagent bottle exists; (5) The gun head is combined with the tip head, and the tip head sensing component is used to sense whether the gun head is successfully combined with the tip head.

10. The method according to claim 9, wherein Step (1) also includes taking a photo of the upper surface of the nucleic acid extraction and amplification box through a camera component, comparing it with the stored information, and judging whether the tip head and the reagent bottle are placed correctly. If it is found that the order of the reagent bottles is placed incorrectly, the stored placement information of the reagent bottles and tools in the nucleic acid extraction and amplification box is adjusted, and the operation is performed according to the actual placement order.

Images