An automatic nucleic acid detection apparatus and a nucleic acid detection method thereof

Abstract

The application relates to an automatic nucleic acid detection device and a nucleic acid detection method thereof. The automatic nucleic acid detection device comprises a fixing frame, a cartridge positioning assembly with a cartridge loading position, a lifting assembly comprising a lifting seat capable of moving controllably close to or away from the cartridge loading position, a rotating assembly comprising a rotating member arranged on the lifting seat, the rotating member being capable of rotating controllably relative to the lifting seat around a rotating axis, the rotating axis being parallel to a first direction, and a pipetting assembly mounted on the rotating member and having a gun head connecting portion, the pipetting assembly being capable of rotating with the rotating member to align the gun head connecting portion with any reagent bin of the cartridge in the first direction, the pipetting assembly being capable of moving with the lifting seat to an insertion position or an exit position; when the pipetting assembly moves to the insertion position, a gun head on the gun head connecting portion is inserted into the reagent bin of the cartridge aligned therewith; when the pipetting assembly moves to the exit position, the gun head on the gun head connecting portion is withdrawn from the reagent bin of the cartridge aligned therewith.

Description

Technical Field

[0001] This invention relates to the field of biological detection technology, and in particular to an automated nucleic acid detection device and its nucleic acid detection method. Background Technology

[0002] Nucleic acid detection generally employs PCR (Polymerase Chain Reaction) technology. PCR is a molecular biology technique that amplifies specific DNA (deoxyribonucleic acid) sequences in vitro. Due to its high specificity, high sensitivity, low purity requirements, and simplicity and speed, PCR is widely used in nucleic acid detection and analysis.

[0003] When using PCR technology for nucleic acid detection, nucleic acid extraction and pre-detection processing of samples are required. However, the processes of nucleic acid extraction and pre-detection processing are quite complex, requiring sample or reagent transfer at each step. This complexity results in low automation levels in the equipment, necessitating manual assistance, and also leads to complex and bulky equipment requiring significant space. For example, a fully automated nucleic acid extraction and analysis integrated device disclosed in patent application number 202110852301.7 uses a first electrode and a lead screw structure to drive the movement of the test cartridge relative to the reagent capsule holder, and a sixth electrode to drive the up-and-down movement of the injection column via belt drive and a lead screw structure. This complex driving structure contributes to the overall complexity and bulkiness of the device. Summary of the Invention

[0004] Therefore, it is necessary to address the problems of existing nucleic acid detection equipment, such as complex structure, large size, large space requirement, and low degree of automation, by providing an automated nucleic acid detection device and its nucleic acid detection method that improves upon the above-mentioned defects.

[0005] An automated nucleic acid testing device includes:

[0006] Fixture;

[0007] A card box positioning assembly is mounted on the fixing frame and has a card box loading position for loading card boxes;

[0008] The lifting assembly includes a lifting seat disposed on the fixed frame, the lifting seat being configured to move controllably toward or away from the card loading position in a first direction;

[0009] A rotating assembly includes a rotating member disposed on the lifting seat, the rotating member being controllably rotatable relative to the lifting seat about a rotation axis, the rotation axis being parallel to the first direction; and

[0010] A pipette assembly is mounted on the rotating member and has a tip connection portion for connecting a tip; the pipette assembly is rotatable with the rotating member to align the tip connection portion with any reagent reservoir of the cartridge in the first direction; the pipette assembly is movable with the lifting seat to an insertion position or an exit position; when the pipette assembly is moved to the insertion position, a tip on the tip connection portion of the pipette assembly is inserted into the reagent reservoir of the cartridge aligned therewith; when the pipette assembly is moved to the exit position, a tip on the tip connection portion of the pipette assembly is withdrawn from the reagent reservoir of the cartridge aligned therewith.

[0011] In one of the embodiments, the rotating assembly further comprises a poking lever, which is eccentrically mounted on the rotating member on a side facing the cartridge loading position;

[0012] The poking lever is movable with the lifting seat along the first direction to an insertion position or an exit position of the cartridge; when the poking lever is inserted into the cartridge, the poking lever can drive the cartridge to rotate with the rotating member.

[0013] In one of the embodiments, the pipette assembly is further rotatable with the rotating member to align the tip connection portion with a tip cavity of the cartridge in the first direction, and when the pipette assembly is moved with the lifting seat to the insertion position, the tip connection portion is connectable with or separable from a tip in the tip cavity of the cartridge.

[0014] In one of the embodiments, the lifting seat sequentially passes through a first position, a second position and a third position during the approaching movement towards the cartridge;

[0015] When the lifting seat is at the first position, the poking lever and the tip connection portion are both withdrawn from the cartridge; when the lifting seat is at the second position, the pipette assembly is at the exit position and the poking lever is inserted into the cartridge; when the lifting seat is at the third position, the pipette assembly is at the insertion position.

[0016] In one of the embodiments, the pipette assembly is rotatable with the rotating member to align the tip connection portion with a plunger hole on the cartridge in the first direction, and is movable with the lifting seat along the first direction to approach the cartridge to insert into the plunger hole on the cartridge and push the plunger.

[0017] In one of the embodiments, the lifting assembly further comprises a driving mechanism, which is drivingly connected between the fixed frame and the lifting seat to drive the lifting seat to move along the first direction relative to the fixed frame.

[0018] In one of the embodiments, the driving mechanism comprises a linear driving member, which is mounted on the fixed frame and drivingly connected with the lifting seat to drive the lifting seat to move relative to the fixed frame along the first direction; or

[0019] The driving mechanism comprises a lead screw, a lifting driving member and a lead screw nut; the lead screw is rotatably connected with the fixed frame along its axis, and the axis of the lead screw is parallel to the first direction; the lifting driving member is mounted on the fixed frame and drivingly connected with the lead screw; the lead screw nut is threadedly connected with the lead screw and fixedly connected with the lifting seat.

[0020] In one of the embodiments, the lifting assembly further comprises a guide rod, which is fixedly connected with the fixed frame and parallel to the first direction, and the lifting seat is slidingly connected with the guide rod;

[0021] The lifting seat is provided with an inductive sheet, and the fixed frame is further mounted with two second photoelectric sensors for detecting the inductive sheet passing by, and the two second photoelectric sensors are spaced apart along the first direction; during the movement of the lifting seat along the first direction, the inductive sheet is driven to move between the two second photoelectric sensors.

[0022] In one of the embodiments, the rotating assembly further comprises a rotating driving mechanism, which comprises a rotating driving member, a driving pulley, a driven pulley and a transmission belt;

[0023] The rotating driving member is mounted on the fixed frame, the driving pulley is drivingly connected with the output shaft of the rotating driving member, the driven pulley is mounted on the rotating member and synchronously rotates with the rotating member, and the transmission belt is sleeved between the driving pulley and the driven pulley.

[0024] In one of the embodiments, the automatic nucleic acid detection device further comprises a nucleic acid detection module mounted on the fixed frame, which is arranged corresponding to the cartridge and used for detecting the nucleic acid in the nucleic acid extraction solution formed in the cartridge.

[0025] A nucleic acid detection method using the automatic nucleic acid detection device according to any one of the above embodiments, comprising the following steps:

[0026] a. loading the cartridge on the cartridge loading position of the cartridge positioning assembly; wherein each reagent compartment of the cartridge is respectively preloaded with a sample, various reagents for nucleic acid extraction and pretreatment before detection;

[0027] b. rotating the pipetting assembly by the rotating member until the gun head connecting portion of the pipetting assembly is aligned with any reagent compartment of the cartridge. b. rotating the pipetting assembly by the rotating member until the gun head connecting portion of the pipetting assembly is aligned with any reagent compartment of the cartridge.

[0028] c. the pipetting assembly is driven by the lifting seat to insert or withdraw the tip on the tip connection portion into or out of the aligned reagent cartridge, and when the tip on the tip connection portion is inserted into the aligned reagent cartridge, the pipetting assembly sucks or injects the reagent through the tip on the tip connection portion;

[0029] d. steps b and c are cyclically executed to realize the transfer and mixing of the reagent in each reagent cartridge of the cartridge with the sample, until the nucleic acid extraction and detection pretreatment is completed, and the liquid containing nucleic acid is obtained;

[0030] e. the nucleic acid detection is performed on the liquid containing nucleic acid.

[0031] The automatic nucleic acid detection device and the nucleic acid detection method thereof described above, initially, the sample, the various reagents for nucleic acid extraction and for detection pretreatment are respectively preloaded into each reagent cartridge of the cartridge, and the cartridge is positioned on the cartridge loading position of the cartridge positioning assembly.

[0032] Firstly, the pipetting assembly is rotated to the tip connection portion aligned with one reagent cartridge in the first direction by controlling the rotation of the rotating member. Then, the pipetting assembly is moved to the tip on the tip connection portion inserted into the aligned reagent cartridge (i.e. insertion position) and the reagent in the reagent cartridge is sucked by controlling the lifting seat to move towards the cartridge in the first direction X. After the reagent is sucked, the lifting seat is controlled to move away from the cartridge in the first direction, so that the tip on the tip connection portion of the pipetting assembly is withdrawn from the reagent cartridge (i.e. withdrawal position). Then, the pipetting assembly is rotated to the tip connection portion aligned with the reagent cartridge preloaded with the sample in the first direction by controlling the rotation of the rotating member. Then, the pipetting assembly is moved to the tip on the tip connection portion inserted into the aligned reagent cartridge (i.e. insertion position), and the sucked reagent is injected into the current reagent cartridge to mix the reagent with the sample by controlling the lifting seat to move towards the cartridge in the first direction. After the reagent is injected, the lifting seat is controlled to move away from the cartridge in the first direction, so that the tip on the tip connection portion is withdrawn from the reagent cartridge (i.e. withdrawal position). Then, in the same way as described above, the sample is mixed with other reagents, the extraction and detection pretreatment of nucleic acid are realized, and the liquid containing nucleic acid is obtained, so as to perform nucleic acid detection on the liquid containing nucleic acid.

[0033] Thus, the lifting seat drives the rotating component and pipetting assembly to move along the first direction to insert or withdraw reagents from the reagent compartment, achieving reagent aspiration or injection. The rotating component drives the pipetting assembly to rotate, allowing the pipette tip on the tip connector to be selectively aligned with any reagent compartment for easy reagent transfer. In other words, nucleic acid extraction and pre-detection treatment only require driving the pipetting assembly to move along the first direction and rotate around the axis of rotation, eliminating the need for a drive structure to move and / or rotate the cartridges, simplifying the equipment structure and reducing its size. Furthermore, nucleic acid extraction, pre-detection treatment, and nucleic acid detection are all automated, with a high degree of automation, requiring no manual intervention, reducing experimental errors. Reagent mixing is completed within the cartridge, reducing aerosol contamination and hazards to laboratory personnel. Attached Figure Description

[0034] Figure 1 This is a front view of an automated nucleic acid testing device according to an embodiment of the present invention;

[0035] Figure 2 for Figure 1 A 3D view of the automated nucleic acid testing equipment shown;

[0036] Figure 3 for Figure 1 The diagram shows the structure of the cartridge positioning component and the nucleic acid detection module of the automated nucleic acid testing device.

[0037] Figure 4 for Figure 1 The diagram shows the structure of the rotating and lifting components of the automated nucleic acid testing equipment.

[0038] Figure 5 This is a flowchart of the steps of a nucleic acid detection method in one embodiment of the present invention;

[0039] Figure 6 This is a flowchart illustrating the steps of picking up the pipette tip in a nucleic acid detection method according to an embodiment of the present invention.

[0040] Figure 7 This is a flowchart illustrating the steps of injecting a plunger in a nucleic acid detection method according to an embodiment of the present invention. Detailed Implementation

[0041] To make the above-mentioned objects, features, and advantages of the present invention more apparent and understandable, specific embodiments of the present invention will be described in detail below with reference to the accompanying drawings. Many specific details are set forth in the following description to provide a thorough understanding of the present invention. However, the present invention can be practiced in many other ways different from those described herein, and those skilled in the art can make similar modifications without departing from the spirit of the present invention. Therefore, the present invention is not limited to the specific embodiments disclosed below.

[0042] In the description of the application, it should be understood that the orientation or positional relationship indicated by the terms "center", "longitudinal", "lateral", "length", "width", "thickness", "upper", "lower", "front", "back", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", "clockwise", "counterclockwise", "axial", "radial", "circumferential" and the like is based on the orientation or positional relationship shown in the drawings, and is only for the convenience of describing the application and simplifying the description, and does not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore cannot be understood as a limitation on the application.

[0043] In addition, the terms "first", "second" are only for descriptive purposes and cannot be understood as indicating or implying relative importance or implicitly indicating the number of the technical features indicated. Therefore, the features defined with "first", "second" can explicitly or implicitly include at least one of the features. In the description of the application, the meaning of "a plurality of" is at least two, such as two, three, etc., unless otherwise explicitly specified and limited.

[0044] In the present application, unless otherwise explicitly specified and limited, the terms "mounting", "connection", "connection", "fixing" and the like should be understood broadly, for example, it can be fixed connection, or detachable connection, or integral; it can be mechanical connection, or electrical connection; it can be directly connected, or indirectly connected through intermediate medium, it can be the internal communication of two elements or the interaction relationship between two elements, unless otherwise explicitly limited. For those skilled in the art, the specific meaning of the above terms in the present application can be understood according to the specific circumstances.

[0045] In the present application, unless otherwise explicitly specified and limited, the first feature is "on" or "under" the second feature, which can be direct contact between the first and second features, or indirect contact between the first and second features through intermediate medium. Moreover, the first feature "above", "above" and "above" the second feature can be directly above or obliquely above the first feature, or only indicate that the horizontal height of the first feature is higher than that of the second feature. The first feature "below", "below" and "below" the second feature can be directly below or obliquely below the first feature, or only indicate that the horizontal height of the first feature is less than that of the second feature.

[0046] It is to be understood that when an element as a preamble is referred to as being "on" or "connected" to another element, it can be directly on or connected to the other element or intervening elements can also be present. As used herein, the terms "vertical", "horizontal", "upper", "lower", "left", "right", and the like as well as the like terms are used for description only and not of the only embodiments.

[0047] Referring to Figure 1 An embodiment of the present application provides an automatic nucleic acid detection device, comprising a fixing frame 10, a cartridge positioning assembly 20, a lifting assembly 30, a rotating assembly 40 and a pipetting assembly 50.

[0048] The cartridge positioning assembly 20 is mounted on the fixing frame 10 and has a cartridge loading position (not labeled in the figure) for loading a cartridge 100. The lifting assembly 30 comprises a lifting seat 31 arranged on the fixing frame 10. The lifting seat 31 is configured to be controllably moved towards or away from the cartridge loading position along a first direction X. The rotating assembly 40 comprises a rotating member 41 arranged on the lifting seat 31 and controllably rotatable relative to the lifting seat 31 about a rotating axis parallel to the first direction X.

[0049] The pipetting assembly 50 is mounted on the rotating member 41, so that the pipetting assembly 50 can rotate with the rotating member 41, and the rotating member 41 and the pipetting assembly 50 can move along the first direction X with the lifting seat 31. The pipetting assembly 50 has a tip connecting portion 51 for connecting a tip, and the pipetting assembly 50 can be rotated with the rotating member 41 to align the tip connecting portion 51 with any reagent chamber (not shown in the figure) of the cartridge 100 in the first direction X. The pipetting assembly 50 can be moved along the first direction X with the lifting seat 31 to an insertion position or an exit position. When the pipetting assembly 50 is moved to the insertion position, a tip 106 (see Figure 3 ) on the tip connecting portion 51 is inserted into the reagent chamber of the cartridge 100 aligned therewith to suck or inject reagents. When the pipetting assembly 50 is moved to the exit position, the tip 106 on the tip connecting portion 51 is withdrawn from the reagent chamber of the cartridge 100 corresponding thereto.

[0050] The above-mentioned automatic nucleic acid detection device initially preloads samples, various reagents for nucleic acid extraction and pre-treatment for detection into respective reagent chambers of the cartridge 100, and positions the cartridge 100 on the cartridge loading position of the cartridge positioning assembly 20.

[0051] Firstly, the rotating member 41 is controlled to rotate, so that the pipetting assembly 50 rotates with the rotating member 41 to align the gun head connecting portion 51 with one reagent cartridge in the first direction X. Then, the lifting seat 31 is controlled to move towards the cartridge 100 along the first direction X, so that the gun head 106 on the gun head connecting portion 51 is inserted into the reagent cartridge aligned therewith (i.e. insertion position), and the reagent in the reagent cartridge is aspirated. After the reagent is aspirated, the lifting seat 31 is controlled to move away from the cartridge 100 along the first direction X, so that the gun head 106 on the gun head connecting portion 51 is withdrawn from the reagent cartridge (i.e. withdrawal position). Then, the rotating member 41 is controlled to rotate, so that the pipetting assembly 50 rotates with the rotating member 41 to align the gun head connecting portion 51 with the reagent cartridge preloaded with a sample in the first direction X. Then, the lifting seat 31 is controlled to move towards the cartridge 100 along the first direction X, so that the gun head 106 on the gun head connecting portion 51 is inserted into the reagent cartridge aligned therewith (i.e. insertion position), and the aspirated reagent is injected into the reagent cartridge, so that the reagent is mixed with the sample. After the reagent is injected, the lifting seat 31 is controlled to move away from the cartridge 100 along the first direction X, so that the gun head 106 on the gun head connecting portion 51 is withdrawn from the reagent cartridge (i.e. withdrawal position). Then, in the same way as described above, the sample is mixed with other reagents, the extraction and pre-treatment of nucleic acid are realized, and a liquid containing nucleic acid is obtained, so that the nucleic acid detection is performed on the liquid containing nucleic acid.

[0052] In this way, the lifting seat 31 drives the rotating member 41 and the pipetting assembly 50 to move along the first direction X to insert or withdraw the reagent cartridge, so that the reagent is aspirated or injected. The rotating member 41 drives the pipetting assembly 50 to rotate, so that the gun head 106 on the gun head connecting portion 51 can be selectively aligned with any reagent cartridge, so as to realize the transfer of the reagent. That is, only the pipetting assembly 50 needs to be driven to move along the first direction X and rotate around the rotation axis, so that the extraction and pre-treatment of nucleic acid are realized, without the need to set a driving structure for driving the cartridge 100 to move and / or rotate, which is beneficial to simplify the structure of the device and reduce the size of the device. Moreover, the extraction and pre-treatment of nucleic acid and the nucleic acid detection can be realized automatically, with a high degree of automation, without the need for manual intervention of personnel, which reduces the error of the test and detection, and the mixing of reagents is completed in the cartridge, which reduces the pollution of aerosol and the harm to the experimental personnel.

[0053] In the embodiment, the automatic nucleic acid detection device further comprises a nucleic acid detection module 60 arranged on the fixed frame 10. The nucleic acid detection module 60 is arranged corresponding to the cartridge 100, so as to perform nucleic acid detection (e.g. fluorescence detection) on the liquid containing nucleic acid obtained by the extraction and pre-treatment of nucleic acid in the cartridge 100.

[0054] Please refer to Figure 3As shown, it should be noted that the cartridge 100 comprises a cartridge body 101, an upper cover 102 and a PCR tube 103, each reagent bin is opened on the cartridge body 101, and the cartridge body 101 can be loaded on the cartridge loading position of the cartridge positioning assembly 20. The upper cover 102 is arranged on the cartridge body 101 and can rotate relative to the cartridge body 101. The upper cover 102 is provided with a gun head cavity 105 and a poking hole 104, and the upper cover 102 can be rotated to be aligned with any reagent bin on the cartridge body 101 in a first direction. The gun head cavity 105 is used to accommodate a gun head 106, and when the gun head 106 is accommodated in the gun head cavity 105, the head of the gun head 106 extends into the reagent bin aligned with the gun head cavity 105. The poking hole 104 is used for the insertion of the following poking rod 42.

[0055] Please refer to Figures 1 to 3 As shown, in the embodiment of the present application, the rotating assembly 40 further comprises a poking rod 42, which is eccentrically mounted on the side of the rotating piece 41 facing the cartridge loading position, so that the poking rod 42 can rotate with the rotating piece 41 and can also move along with the lifting seat 31 in the first direction X. The poking rod 42 can move along with the lifting seat 31 in the first direction X to a position where the poking rod 42 is inserted into or withdrawn from the poking hole 104 on the upper cover 102 of the cartridge 100. When the poking rod 42 is inserted into the poking hole 104 on the upper cover 102 of the cartridge 100, the poking rod 42 can drive the upper cover 102 to rotate with the rotating piece 41 (the cartridge body 101 is fixed and does not rotate with the upper cover 102). In this way, the gun head on the gun head connecting part 51 can be aligned with any reagent bin in the first direction by driving the upper cover 102 to rotate with the rotating piece 41 by the poking rod 42, so as to facilitate the transfer of reagents.

[0056] In specific embodiments, the pipetting assembly 50 can also rotate with the rotating piece 41 to align the gun head connecting part 51 with the gun head cavity 105 of the cartridge 100 in the first direction (X).

[0057] When the gun head connecting part 51 is aligned with the gun head cavity 105 of the cartridge 100 in the first direction (X) and the pipetting assembly 50 moves with the lifting seat 31 to the insertion position, the gun head connecting part 51 can be connected or separated from the gun head 106 in the gun head cavity 105. In this way, when it is necessary to pick up the gun head 106, the pipetting assembly 50 moves with the lifting seat 31 to the insertion position, so that the gun head connecting part 51 is inserted into the gun head cavity 105 of the upper cover 102 and presses against the gun head 106 in the gun head cavity 105, so that the gun head 106 is connected with the gun head connecting part 51, that is, the gun head 106 is picked up. When it is necessary to release the gun head 106, the pipetting assembly 50 moves with the lifting seat 31 to the insertion position, so that the gun head connecting part 51 drives the gun head 106 to be inserted into the gun head cavity 105 of the upper cover 102 and presses against the gun head 106, so that the gun head 106 is disconnected from the gun head connecting part 51, that is, the gun head 106 is released into the gun head cavity 105 of the upper cover 102.

[0058] In actual operation, when the reagent in a selected reagent cartridge is to be aspirated or the aspirated reagent is to be injected into the selected reagent cartridge, the lifting seat 31 moves towards the cartridge 100 along the first direction X until the poking rod 42 is inserted into the poking hole 104 on the upper cover 102 and the gun head connecting portion 51 of the pipette assembly 50 is inserted into the gun head cavity 105 on the upper cover 102 and connected with the gun head 106. The control rotating member 41 is rotated so that the poking rod 42 pokes the upper cover 102 to rotate relative to the cartridge body 101 until the gun head cavity 105 is aligned with the selected reagent cartridge. Then, the lifting seat 31 is controlled to move towards the cartridge 100 along the first direction X so that the pipette assembly 50 drives the gun head 106 to be inserted into the selected reagent cartridge, thereby aspirating the reagent in the selected reagent cartridge or injecting the reagent into the selected reagent cartridge.

[0059] In specific embodiments, the lifting seat 31 sequentially passes through the first position, the second position and the third position during the process of moving towards the cartridge 100, that is, the third position is closest to the cartridge 100, the first position is farthest from the cartridge 100, and the second position is between the first position and the third position. When the lifting seat 31 moves to the first position, the poking rod 42 and the gun head connecting portion 51 of the pipette assembly 50 are both withdrawn from the cartridge 100, that is, the poking rod 42 is withdrawn from the poking hole 104 on the upper cover 102 and the gun head connecting portion 51 is withdrawn from the gun head cavity 105 on the upper cover 102. When the lifting seat 31 moves to the second position, the poking rod 42 is inserted into the cartridge 100, and the pipette assembly 50 is located at the above-mentioned withdrawn position, that is, the gun head 106 on the gun head connecting portion 51 is inserted into the gun head cavity 105 on the upper cover 102, and the gun head 106 on the gun head connecting portion 51 is not inserted into the current reagent cartridge, at this time, the gun head 106 will not interfere with the rotational movement of the upper cover 102 relative to the cartridge body 101. When the lifting seat 31 moves to the third position, the pipette assembly 50 is located at the above-mentioned inserted position, that is, the gun head 106 on the gun head connecting portion 51 is inserted into the current reagent cartridge from the gun head cavity 105, at this time, the reagent can be injected into the reagent cartridge or the reagent in the reagent cartridge can be aspirated.

[0060] When it is necessary to inject reagent into a selected reagent chamber or to suck reagent in a selected reagent chamber, first, the lifting seat 31 moves from the first position to the third position along the first direction X, until the poking rod 42 is inserted into the poking hole 104 of the upper cover 102 (the insertion depth is deep), and the gun head connecting portion 51 of the pipetting assembly 50 is inserted into the gun head cavity 105 and presses against the gun head 106, so that the gun head connecting portion 51 is connected with the gun head 106. Then, the lifting seat 31 moves from the third position to the second position along the first direction X away from the cartridge 100. At this time, the poking rod 42 remains inserted into the poking hole 104 of the upper cover 102 (the insertion depth is shallow), and the gun head 106 on the gun head connecting portion 51 is withdrawn from the reagent chamber (at this time, the gun head 106 is only in the gun head cavity 105, so that the gun head 106 does not interfere with the rotation of the upper cover 102 relative to the cartridge body 101). Then, the rotating member 41 rotates, so that the poking rod 42 rotates the upper cover 102, and at the same time, the pipetting assembly 50 also rotates with the rotating member 41, until the gun head 106 on the pipetting assembly 50 is aligned with the selected reagent chamber in the first direction X. Then, the lifting seat 31 moves from the second position to the third position along the first direction X towards the cartridge 100, until the pipetting assembly 50 inserts the gun head 106 on the gun head connecting portion 51 into the selected reagent chamber, at which time the pipetting assembly 50 sucks reagent in the selected reagent chamber through the gun head 106 or injects reagent into the selected reagent chamber.

[0061] It should be noted that one of the reagent chambers of the cartridge body 101 is in communication with the detection chamber of the PCR tube 103 (for ease of description, the reagent chamber is named as the injection chamber), and the upper cover 102 has a plunger hole 107 in which a plunger (not shown in the figure) is arranged, so that the plunger can be used to inject the liquid containing nucleic acid in the injection chamber into the detection chamber of the PCR tube 103, so as to facilitate subsequent nucleic acid detection of the detection chamber of the PCR tube 103.

[0062] In specific embodiments, the pipetting assembly 50 can also rotate with the rotating member 41 to a position where the gun head connecting portion 51 is aligned with the plunger hole 107 on the cartridge 100 in the first direction X, and can be inserted into the plunger hole 107 on the cartridge 100 and push the plunger to move when the lifting seat 31 moves along the first direction X towards the cartridge 100. In this way, the gun head connecting portion 51 pushes the plunger to move from the plunger hole 107 into the injection chamber aligned therewith, so as to inject the liquid containing nucleic acid in the injection chamber into the detection chamber of the PCR tube 103.

[0063] Thus, after nucleic acid extraction and pre-detection processing are completed, and the nucleic acid-containing liquid is transferred into the injection chamber, the lifting seat 31 moves along the first direction X towards the cartridge 100 until it reaches the third position, allowing the pipetting assembly 50 to insert the pipette tip 106 into the corresponding reagent compartment and press against the pipette tip 106, causing the pipette tip 106 to disengage from the pipette tip connection part 51 of the pipetting assembly 50. Then, the lifting seat 31 moves back to the first position, causing both the pipetting assembly 50 and the actuating lever 42 to exit the cartridge 100. Next, the rotating component 41 is controlled to rotate, thereby driving the actuating lever 42 to rotate to a position aligned with another actuating hole 104 in the first direction X, at which point the pipette tip connection part 51 of the pipetting assembly 50 is aligned with the plunger hole 107. Then, the lifting seat 31 is controlled to move towards the cartridge 100 until the actuating lever 42 is inserted into the corresponding actuating hole 104 and the pipette tip connection part 51 is inserted into the plunger hole 107. Next, the rotating component 41 rotates, causing the actuating lever 42 to rotate the upper cover 102 until the plunger hole 107 is aligned with the injection chamber in the first direction X. Then, the lifting seat 31 moves closer to the cartridge 100 along the first direction X, causing the nozzle connection 51 to push the plunger into the injection chamber until the nucleic acid-containing liquid in the injection chamber is injected into the detection chamber of the PCR tube 103.

[0064] In a specific embodiment, there are two levers 42. The two levers 42 simultaneously rotate the upper cover 102 relative to the card holder body 101, making the rotation of the upper cover 102 more stable and reliable. Optionally, the two levers 42 are symmetrically arranged relative to the rotation axis of the rotating component 41.

[0065] Please see Figure 1 , Figure 2 and Figure 4 As shown in the embodiment of the present invention, the lifting assembly 30 further includes a drive mechanism (not shown), which is tractively connected between the fixed frame 10 and the lifting seat 31 to drive the lifting seat 31 to move relative to the fixed frame 10 along the first direction X. Thus, the lifting seat 31 is driven to move closer to or further away from the card holder 100 along the first direction X by the drive mechanism.

[0066] In one embodiment, the drive mechanism includes a linear drive 32, which is mounted on the fixed frame 10 and drivenly connected to the lifting seat 31 to drive the lifting seat 31 to move relative to the fixed frame 10 along a first direction X. Optionally, the linear drive 32 may be a linear motor or a linear drive module, etc., and is not limited thereto.

[0067] In another embodiment, the driving mechanism comprises a screw rod, a lifting driving member and a screw nut. The screw rod is rotatably connected to the fixed frame 10 about its own axis, and the axis of the screw rod is parallel to the first direction X. The lifting driving member is installed on the fixed frame 10 and is in driving connection with the screw rod to drive the screw rod to rotate about its own axis. The screw nut is in threaded connection with the screw rod and is fixedly connected with the lifting seat 31, so that when the screw rod rotates about its own axis, the screw nut is driven to move along the axis of the screw rod (i.e. along the first direction X), thereby driving the lifting seat 31 to move along the first direction X. Optionally, the lifting driving member can be a motor.

[0068] In a specific embodiment, the lifting assembly 30 further comprises a guide rod 35 fixedly connected to the fixed frame 10 and parallel to the first direction X. The lifting seat 31 is in sliding connection with the guide rod 35, so that the movement of the lifting seat 31 along the first direction X is guided by the guide rod 35, making the movement of the lifting seat 31 more stable and reliable. Optionally, both ends of the guide rod 35 are fixedly connected to the fixed frame 10, and the guide rod 35 is assembled with the lifting seat 31 through a linear bearing 36.

[0069] Further, the guide rod 35 can comprise at least two guide rods 35, which simultaneously guide the lifting seat 31, thereby improving the guiding effect.

[0070] In an embodiment of the present application, the rotating assembly 40 further comprises a rotating driving mechanism (not marked in the figure), which comprises a rotating driving member 48, a driving pulley 43, a driven pulley 44 and a transmission belt 45.

[0071] The rotating driving member 48 is installed on the fixed frame 10, and the driving pulley 43 is in driving connection with the output shaft of the rotating driving member 48, so that the rotating driving member 48 can drive the driving pulley 43 to rotate. The driven pulley 44 is installed on the rotating member 41 and can rotate synchronously with the rotating member 41. The transmission belt 45 is sleeved between the driving pulley 43 and the driven pulley 44, so that when the rotating member 41 drives the driving pulley 43 to rotate, the driven pulley 44 can be driven to rotate through the transmission belt 45, and in turn the rotating member 41 is driven to rotate by the driven pulley 44. Preferably, the rotating driving member 48 can be a motor. The driving pulley 43 and the driven pulley 44 can be synchronous pulleys, and the transmission belt 45 can be a synchronous belt.

[0072] In a specific embodiment, the rotating member 41 is a cylindrical member, and the lifting seat 31 has a mounting hole (not shown in the figure), and the cylindrical member is sleeved in the mounting hole through a bearing, so that the cylindrical member can rotate relative to the lifting seat 31. The driven pulley 44 can be coaxially installed on the cylindrical member through threaded fasteners (such as bolts, etc.), so that the driven pulley 44 can drive the rotating member 41 to rotate synchronously.

[0073] Further, the toggle lever 42 is fixedly connected to one end of the cylinder member facing the cartridge 100. The pipette assembly 50 is fixedly installed in the inner cavity of the cylinder member, and the gun head connecting portion 51 penetrates through the one end of the cylinder member facing the cartridge 100, so as to be connected or separated with the gun head 106.

[0074] In the embodiment, the rotating member 41 is provided with a code disc 46, and the code disc 46 is provided with marks for identifying the positions of the reagent chambers of the cartridge 100. The fixed frame 10 is provided with a first photoelectric sensor 47, and the first photoelectric sensor 47 is used to detect the marks on the code disc 46, so as to accurately rotate the upper cover 102 of the cartridge 100 to the required position, and then accurately inject reagents into the reagent chambers or suck the reagents in the reagent chambers.

[0075] In the embodiment, the lifting seat 31 is provided with the inductive sheet 34, and the fixed frame 10 is further provided with two second photoelectric sensors 33, which are arranged along the first direction X and are used to detect the inductive sheet 34. During the movement of the lifting seat 31 along the first direction X, the inductive sheet 34 is driven to move between the two second photoelectric sensors 33, so as to limit the maximum stroke of the lifting seat 31 moving along the first direction X by the two second photoelectric sensors 33.

[0076] Based on the automatic nucleic acid detection device, the application further provides a nucleic acid detection method using the automatic nucleic acid detection device.

[0077] S10, loading the cartridge 100 on the cartridge loading position of the cartridge positioning assembly 20. Each reagent chamber of the cartridge 100 is preloaded with a sample, various reagents for nucleic acid extraction and pretreatment before detection.

[0078] S20, rotating the pipette assembly 50 by the rotating member 41 until the gun head connecting portion 51 of the pipette assembly 50 is aligned with any reagent chamber of the cartridge 100. Specifically, the lifting seat 31 is located at the second position, at this time, the toggle lever 42 is inserted into the toggle hole 104, and the gun head 106 on the gun head connecting portion 51 of the pipette assembly 50 is located in the gun head cavity 105. The rotating member 41 drives the toggle lever 42 and the pipette assembly 50 to rotate, so that the toggle lever 42 rotates the upper cover 102 of the cartridge 100, until the gun head connecting portion 51 of the pipette assembly 50 is aligned with any reagent chamber of the cartridge 100.

[0079] S30, inserting or withdrawing the gun head 106 on the gun head connecting portion 51 into or out of the current reagent chamber by the lifting seat 31 driving the pipette assembly 50, and when the gun head 106 on the gun head connecting portion 51 is inserted into the current reagent chamber, the pipette assembly 50 sucks the reagent in the current reagent chamber or injects the reagent into the current reagent chamber through the gun head 106.

[0080] Specifically, the lifting seat 31 is moved from the second position to the third position, so that the gun head 106 on the gun head connecting portion 51 of the pipette assembly 50 is inserted into the current reagent cartridge, at this time the pipette assembly 50 injects or sucks the reagent in the current reagent cartridge through the gun head 106. Then, the lifting seat 31 is returned from the third position to the second position, so that the tumbler rod 42 remains inserted into the tumbler hole 104, the gun head 106 on the gun head connecting portion 51 exits the current reagent cartridge and is located in the gun head cavity 105, so that the tumbler rod 42 can tumbler the upper cover 102 to rotate in the subsequent steps, and the gun head 106 does not interfere with the rotation of the upper cover 102 relative to the cartridge body 101.

[0081] S40, steps S20 and S30 are repeatedly executed to realize the transfer and mixing of the reagent in each reagent cartridge and the sample until the nucleic acid extraction and detection pretreatment is completed, and the liquid containing nucleic acid is obtained.

[0082] S50, nucleic acid detection is performed on the liquid containing nucleic acid. Specifically, the nucleic acid detection module 60 is used to perform nucleic acid detection on the liquid containing nucleic acid in the detection chamber of the PCR tube 103.

[0083] Specific to the embodiment, before step S20, it further includes the step of picking up the gun head 106:

[0084] S201, the lifting seat 31 is moved along the first direction X to the third position, so that the tumbler rod 42 is inserted into the tumbler hole 104, the gun head connecting portion 51 of the pipette assembly 50 is inserted into the gun head cavity 105, and the gun head 106 is pressed to connect the gun head 106 and the gun head connecting portion 51;

[0085] S202, the lifting seat 31 is moved along the first direction X to the second position, so that the tumbler rod 42 remains inserted into the tumbler hole 104 (only the insertion depth is shallower), the pipette assembly 50 drives the gun head 106 on the gun head connecting portion 51 to exit the current reagent cartridge and be located in the gun head cavity 105, so that when step S20 is performed, the tumbler rod 42 can tumbler the upper cover 102 to rotate, and the gun head 106 does not interfere with the rotation of the upper cover 102 relative to the cartridge body 101.

[0086] Specific to the embodiment, between steps S40 and S50, it further includes the step of pushing the plunger injection:

[0087] S401, the lifting seat 31 is moved along the first direction X from the second position to the third position, so that the pipette assembly 50 inserts the gun head 106 into the current reagent cartridge (at this time a part of the gun head 106 is located in the gun head cavity 105, and another part is inserted into the current reagent cartridge), and presses the gun head 106 to separate the gun head 106 and the gun head connecting portion 51 of the pipette assembly 50;

[0088] S402, the lifting seat 31 is moved from the third position to the first position along the first direction X, so that the gun head connecting part 51 of the pipette assembly 50 and the poking rod 42 are both out of the cartridge 100 (i.e. the gun head connecting part 51 of the pipette assembly 50 is out of the gun head cavity 105, and the poking rod 42 is out of the poking hole 104);

[0089] S403, the rotating piece 41 is rotated until the gun head connecting part 51 is aligned with the plunger hole 107 on the upper cover 102 in the first direction X, and the poking rod 42 is aligned with another poking hole 104 on the upper cover 102 in the first direction X;

[0090] S404, the lifting seat 31 is moved along the first direction X to approach the cartridge 100 until the gun head connecting part 51 is inserted into the plunger hole 107;

[0091] S405, the rotating piece 41 is rotated until the poking rod 42 drives the upper cover 102 to rotate to a position where the plunger hole 107 is aligned with the reagent bin containing nucleic acid-containing liquid (i.e. the above-mentioned injection bin);

[0092] S406, the lifting seat 31 is moved along the first direction X to approach the cartridge 100, so that the gun head connecting part 51 pushes the plunger to be inserted into the current reagent bin (i.e. the above-mentioned injection bin), until the nucleic acid-containing liquid in the current reagent bin is injected into the detection bin of the PCR tube 103;

[0093] S407, the lifting seat 31 is moved along the first direction X to move away from the cartridge 100 until the poking rod 42 is out of the poking hole 104, and the gun head connecting part 51 is out of the plunger hole 107.

[0094] It should be noted that the specific steps of the experiment using the above-mentioned automatic nucleic acid detection device are not limited, and different experimental processes can be formulated according to needs. For example, in order to avoid mutual contamination of reagents that do not need to be mixed, a step of replacing the gun head can be set after each reagent transfer.

[0095] The technical features of the above-mentioned embodiments can be combined in any way. In order to make the description simple, all possible combinations of the technical features in the above-mentioned embodiments are not described, but as long as the combinations of the technical features do not exist, they should be considered as the scope of the description.

[0096] The above-mentioned embodiments only express several embodiments of the present application, and the description is more specific and detailed, but it should not be understood as a limitation on the scope of the patent. It should be noted that for ordinary skilled persons in the art, without departing from the concept of the present application, a number of modifications and improvements can be made, which are within the scope of the present application. Therefore, the protection scope of the patent of the present application should be subject to the appended claims.

Claims

1. An automated nucleic acid testing device, characterized in that, include: Fixture (10); Card box positioning assembly (20) is mounted on the fixture (10) and has a card box loading position for loading card boxes (100); The lifting assembly (30) includes a lifting seat (31) disposed on the fixed frame (10), the lifting seat (31) being configured to move controllably toward or away from the card loading position in a first direction (X); The rotating assembly (40) includes a rotating member (41) disposed on the lifting seat (31), the rotating member (41) being controllably rotatable relative to the lifting seat (31) about a rotation axis, the rotation axis being parallel to the first direction (X); and A pipetting assembly (50) is mounted on the rotating member (41) and has a pipette tip connector (51) for connecting pipette tips. The pipetting assembly (50) can rotate with the rotating member (41) until the pipette tip connector (51) is aligned with any reagent compartment of the cartridge (100) in the first direction (X). The pipetting assembly (50) can move with the lifting seat (31) to the insertion position or the withdrawal position. The rotating assembly (40) further includes a lever (42), which is eccentrically mounted on the side of the rotating component (41) facing the card holder loading position. The lever (42) can move along the first direction (X) with the lifting seat (31) to the position of inserting or retracting the card holder (100). When the lever (42) is inserted into the card holder (100), the lever (42) can drive the card holder (100) to rotate together with the rotating component (41). When the pipetting assembly (50) moves to the insertion position, the pipette tip (106) on the tip connector (51) is inserted into the reagent compartment of the cartridge (100) that is aligned with it; when the pipetting assembly (50) moves to the withdrawal position, the pipette tip (106) on the tip connector (51) is withdrawn from the reagent compartment of the cartridge (100) that is aligned with it.

2. The automated nucleic acid testing device according to claim 1, characterized in that, The pipetting assembly (50) can also rotate with the rotating member (41) until the tip connection (51) and the tip cavity (105) of the cartridge (100) are aligned in the first direction (X), and when the pipetting assembly (50) moves with the lifting seat (31) to the insertion position, the tip connection (51) can connect or separate from the tip (106) in the tip cavity (105) of the cartridge (100).

3. The automated nucleic acid testing device according to claim 2, characterized in that, As the lifting seat (31) moves toward the card box (100), it passes through the first position, the second position, and the third position in sequence; When the lifting seat (31) is in the first position, the lever (42) and the pipette tip connector (51) are both disengaged from the cartridge (100); when the lifting seat (31) is in the second position, the pipetting assembly (50) is in the disengaged position and the lever (42) is inserted into the cartridge (100); when the lifting seat (31) is in the third position, the pipetting assembly (50) is in the inserted position.

4. The automated nucleic acid testing device according to claim 2, characterized in that, The pipetting assembly (50) can also rotate with the rotating member (41) to a position where the tip connector (51) and the plunger hole (107) on the cartridge (100) are aligned in the first direction (X), and can move with the lifting seat (31) along the first direction (X) closer to the cartridge (100) to insert into the plunger hole (107) on the cartridge (100) and push the plunger.

5. The automated nucleic acid testing device according to any one of claims 1 to 4, characterized in that, The lifting assembly (30) also includes a drive mechanism; The driving mechanism includes a linear drive element (32), which is mounted on the fixed frame (10) and drivenly connected to the lifting seat (31) to drive the lifting seat (31) to move relative to the fixed frame (10) along the first direction (X); or The driving mechanism includes a lead screw, a lifting drive component, and a lead screw nut; the lead screw is rotatably connected to the fixed frame (10) around its own axis, and the axis of the lead screw is parallel to the first direction (X); the lifting drive component is installed on the fixed frame (10) and is connected to the lead screw in a transmission connection; the lead screw nut is threadedly connected to the lead screw and is fixedly connected to the lifting seat (31).

6. The automated nucleic acid testing device according to claim 5, characterized in that, The lifting assembly (30) further includes a guide rod (35), which is fixedly connected to the fixed frame (10) and parallel to the first direction (X). The lifting seat (31) is slidably connected to the guide rod (35). The lifting seat (31) is provided with a sensing plate (34), and the fixed frame (10) is also equipped with two second photoelectric sensors (33) for detecting the sensing plate (34) passing by. The two second photoelectric sensors (33) are arranged at intervals along the first direction (X). During the movement of the lifting seat (31) along the first direction (X), the sensing plate (34) is driven to move between the two second photoelectric sensors (33).

7. The automated nucleic acid testing device according to any one of claims 1 to 4, characterized in that, The rotating assembly (40) further includes a rotating drive mechanism, which includes a rotating drive component (48), a driving pulley (43), a driven pulley (44), and a transmission belt (45). The rotary drive (48) is mounted on the fixed frame (10), the driving pulley (43) is connected to the output shaft of the rotary drive (48), the driven pulley (44) is mounted on the rotary component (41) and rotates synchronously with the rotary component (41), and the transmission belt (45) is sleeved between the driving pulley (43) and the driven pulley (44).

8. The automated nucleic acid testing device according to any one of claims 1 to 4, characterized in that, The automatic nucleic acid testing equipment also includes a nucleic acid testing module (60) installed on the fixed frame (10). The nucleic acid testing module (60) is set corresponding to the card box (100) and is used to perform nucleic acid testing on the nucleic acid extract formed in the card box (100).

9. A nucleic acid detection method using an automated nucleic acid detection device as described in any one of claims 1 to 8, characterized in that, Includes the following steps: a. The card cartridge (100) is loaded onto the card cartridge loading position of the card cartridge positioning assembly (20); wherein, each reagent compartment of the card cartridge (100) is pre-loaded with samples, various reagents for nucleic acid extraction and pre-detection treatment; b. The rotating component (41) drives the pipetting assembly (50) to rotate until the pipetting assembly (50) tip connection (51) is aligned with one of the reagent compartments of the cartridge (100). c. The lifting seat (31) drives the pipetting assembly (50) to insert or withdraw the pipette tip (106) on the pipette tip connector (51) into or out of the aligned reagent compartment. When the pipette tip (106) on the pipette tip connector (51) is inserted into the aligned reagent compartment, the pipetting assembly (50) draws or injects reagent through the pipette tip (106) on the pipette tip connector (51). d. Repeat steps b and c to transfer reagents in each reagent compartment of the cartridge (100) and mix them with the sample until nucleic acid extraction and pre-detection treatment are completed and a liquid containing nucleic acid is obtained; e. Perform nucleic acid testing on liquids containing nucleic acid.

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