Nucleic acid test kits, nucleic acid testing systems, and sample processing methods

Abstract

This disclosure provides a nucleic acid detection cartridge, a nucleic acid detection system, and a sample processing method. The nucleic acid detection cartridge includes: a cartridge body (2) comprising a compartment (200), the compartment (200) including an outer wall enclosing an internal space and multiple partition walls (218) located within the internal space and extending in a vertical direction (A), the multiple partition walls (218) being configured to divide the internal space into multiple chambers, the multiple chambers including a transfer chamber (201) and multiple receiving chambers, the multiple receiving chambers including a sample chamber (291), multiple reagent chambers, and an extraction chamber (20); a rotary valve (5), rotatably disposed on the lower side of the cartridge body (2), including a first flow channel (50), the first flow channel (50) being configured to selectively connect the transfer chamber (201) to any of the receiving chambers; and a drive unit (6), located within the transfer chamber (201), configured to drive fluid to flow between the transfer chamber (201) and the receiving chambers connected via the rotary valve (5).

Description

Technical Field

[0001] This disclosure relates to the field of nucleic acid detection technology, and in particular to a nucleic acid detection cartridge, a nucleic acid detection system, and a sample processing method. Background Technology

[0002] Nucleic acid testing systems encompass functions such as nucleic acid extraction, amplification, and detection. Among these, nucleic acid extraction is the most complex step, requiring multiple processes including sample lysis, adsorption, washing, and elution.

[0003] In related technologies, the overall structure of nucleic acid testing systems is often quite complex and has low integration. This not only makes the actual operation process very cumbersome, but also limits the adaptability of nucleic acid testing systems to different application scenarios. Summary of the Invention

[0004] The purpose of this disclosure is to provide a nucleic acid detection cartridge, a nucleic acid detection system, and a sample processing method, aiming to improve the integration of the nucleic acid detection system and the efficiency of nucleic acid detection.

[0005] The first aspect of this disclosure provides a nucleic acid testing kit, comprising: The cartridge body includes a compartment, which includes an outer wall forming an internal space and multiple partition walls located in the internal space and extending in a vertical direction. The multiple partition walls are configured to divide the internal space into multiple chambers. The multiple chambers include a transfer chamber and multiple receiving chambers. The multiple receiving chambers include a sample chamber, multiple reagent chambers, and an extraction chamber. The multiple reagent chambers include a lysis chamber pre-stored with lysis buffer, a washing chamber pre-stored with washing buffer, an elution chamber pre-stored with elution buffer, and a lyophilized bulb chamber pre-stored with lyophilized bulbs. An amplification chip, mounted on the cartridge body, includes an amplification cavity; A rotary valve, rotatably disposed on the lower side of the cartridge body, includes a first flow channel configured to selectively connect the transfer chamber to either the receiving chamber or the amplification chamber; and The drive unit, located within the transfer chamber, is configured to drive fluid to flow between the transfer chamber and the receiving chamber, which is connected via the rotary valve.

[0006] In some embodiments of the nucleic acid test kit, the chamber further includes a liquid channel, the amplification chamber is connected to the liquid channel, and the first channel of the rotary valve is configured to selectively connect the transfer chamber and the liquid channel.

[0007] In some embodiments of the nucleic acid test kit, the nucleic acid test kit further includes a base, which is connected to the lower side of the kit body, wherein... The card holder body also includes a rotary valve mating part, which is made of elastic material and integrally formed on the lower side of the compartment body, forming a rotary valve cavity between itself and the base. The rotary valve is disposed within the rotary valve cavity and seals with the rotary valve mating part; and / or The cartridge body also includes a mounting part, which is made of elastic material and integrally formed on the front sidewall of the compartment. The mounting part is provided with a through hole for connecting the first flow channel and the amplification cavity of the amplification chip, and the mounting part is used for sealing and engaging with the amplification chip.

[0008] In some embodiments of the nucleic acid test kit, the valve mating part and / or the mounting part are made of TPE, TPU, or liquid silicone material; and / or

[0009] The chamber body is made of PP, COC, or PC material; and / or

[0010] The valve mating part and the housing part are injection molded in two colors, and / or the mounting part and the housing part are injection molded in two colors; and / or

[0011] The surface of the rotating valve mating part that forms the rotating valve cavity mates with the shape of the rotating valve.

[0012] In some embodiments of the nucleic acid test kit, the base includes a side limiting structure located on the front side of the mounting portion, the side limiting structure forming a slot with the mounting portion, the slot being configured to accommodate a connector located on the rear side of the amplification chip, and the mounting portion being configured to seal with the connector.

[0013] In some embodiments of the nucleic acid test kit, the rear surface of the side limiting structure includes a first inclined surface, which forms an acute angle with the vertical direction and gradually slopes backward from top to bottom. The first inclined surface is configured to cooperate with the shape of a second inclined surface disposed on the front surface of the connector.

[0014] In some embodiments of the nucleic acid testing cartridge, the cartridge body includes multiple drainage holes, which are respectively arranged in a one-to-one correspondence with multiple receiving cavities and are capable of communicating with the first flow channel. The drainage hole includes a narrowing section whose diameter gradually decreases from top to bottom; and / or At least one of the receiving cavities has a bottom wall comprising an inclined bottom surface that slopes downward from the bottom end of the partition wall surrounding the receiving cavity to the drainage hole.

[0015] In some embodiments of the nucleic acid test kit, the plurality of chambers further includes a first air chamber, which is separated from the sample chamber by the partition wall; A first air passage is provided at the top of the partition wall separating the first air chamber and the sample chamber, and the first air passage connects the sample chamber and the first air chamber.

[0016] In some embodiments of the nucleic acid test kit, the plurality of receiving cavities further includes a waste liquid cavity, which is separated from the extraction cavity by the partition wall; A second vent is provided at the top of the partition wall separating the waste liquid chamber and the extraction chamber, and the second vent connects the waste liquid chamber and the extraction chamber.

[0017] In some embodiments of the nucleic acid test kit, the waste liquid chamber is further provided with a partition wall, which divides the waste liquid chamber into a waste liquid sub-chamber and a drying sub-chamber. The drying sub-chamber is connected to the extraction chamber through the second air outlet. The first flow channel is configured to selectively connect the transfer chamber to the waste liquid sub-chamber. A desiccant is placed in the drying sub-chamber.

[0018] In some embodiments of the nucleic acid test kit, the separator sub-wall is disposed on the separator wall separating the waste liquid chamber and the extraction chamber; and / or

[0019] A third vent is provided at the top of the partition wall, and the third vent connects the waste liquid chamber and the drying chamber.

[0020] In some embodiments of the nucleic acid test kit, The plurality of chambers also includes a second air chamber, which is configured to keep the air pressure value in the amplification chamber within a preset air pressure range. The valve further includes a second flow channel, which is configured to connect the amplification chamber and the second gas chamber when the first flow channel connects the transfer chamber and the amplification chamber.

[0021] In some embodiments of the nucleic acid test kit, the chamber further includes a gas flow channel, the amplification chamber is connected to the gas flow channel, and the second flow channel is configured to connect the amplification chamber and the second gas chamber through the gas flow channel when the first flow channel connects the transfer chamber and the amplification chamber.

[0022] In some embodiments of the nucleic acid test kit, the main body of the kit includes a first welding part, which is disposed along the top of the outer wall of the compartment and / or the top of the partition wall; The nucleic acid test kit also includes a cover, which is connected to the upper part of the kit body to close at least a portion of the multiple chambers. The cover includes a second welded portion located at the bottom end of the cover and configured to be embedded and welded together with the first welded portion.

[0023] In some embodiments of the nucleic acid test kit, the nucleic acid test kit further includes a cover, the cover being connected above the kit body to close at least a portion of the plurality of chambers, the cover comprising: The lower cover is sealed to the top of the outer wall of the compartment and the top of the plurality of partition walls, and includes a plurality of lower cover openings that communicate with a portion of the plurality of chambers, the portion of the chambers including the sample chamber and the transfer chamber; A sealing film, connected above the lower cover, includes two sealing film openings respectively opposite to the sample chamber and the transfer chamber, for sealing the lower cover openings corresponding to the portions of the chambers other than the sample chamber and the transfer chamber; and An upper cover, movably connected to the lower cover, has a fastened state covering the lower cover and the cover sealing film, and an open state exposing the lower cover and the cover sealing film. It includes a sealing portion and an upper cover opening. In the fastened state, the sealing portion is opposite to the sample cavity and is configured to block the lower cover opening and the sealing film opening corresponding to the sample cavity. The upper cover opening is opposite to the transfer cavity and the sealing film opening corresponding to the transfer cavity.

[0024] In some embodiments of the nucleic acid detection kit, the nucleic acid detection kit further includes an amplification chip, which is mounted on the main body of the kit and includes an amplification cavity; The first flow channel of the rotary valve is configured to selectively connect the transfer chamber to the amplification chamber; The amplification chip further includes a liquid passage for connecting the first flow channel and the amplification cavity and a gas passage for keeping the gas pressure value in the amplification cavity within a preset gas pressure range. The liquid passage includes a large-diameter section away from the amplification cavity, a small-diameter section closer to the amplification cavity with a diameter smaller than the large-diameter section, and a transition section connecting the large-diameter section and the small-diameter section, where the diameter gradually decreases from the large-diameter section to the small-diameter section.

[0025] In some embodiments of the nucleic acid test kit, the plurality of reagent chambers include a plurality of washing chambers, and the plurality of washing chambers are pre-stored with different washing solutions; and / or

[0026] The plurality of reagent chambers include the plurality of lyophilized bulb chambers.

[0027] In some embodiments of the nucleic acid test kit, the bottom of the sample chamber is provided with a drainage hole, which can communicate with the first flow channel; The sample chamber is provided with a filter element extending in the vertical direction, and the bottom of the filter element is arranged opposite to the drainage hole in the sample chamber in the vertical direction.

[0028] In some embodiments of the nucleic acid test kit, the nucleic acid test kit further includes at least one storage container, which is movably located in one of the reagent chambers along the vertical direction so that it can be in a first position and a second position in the reagent chamber; The cartridge body includes a spike, which is located at the bottom of the reagent chamber corresponding to the storage container. At the first position of the storage container, the spike is spaced apart from the storage container in the vertical direction. At the second position of the storage container, the spike pierces the storage container.

[0029] In some embodiments of the nucleic acid test kit, the storage container includes a sealing portion configured to seal the gap between the storage container and the spike portion.

[0030] In some embodiments of the nucleic acid test kit, the sealing portion includes at least one sealing ring disposed at the bottom of the storage container and arranged circumferentially around the spike portion.

[0031] In some embodiments of the nucleic acid test kit, the nucleic acid test kit further includes a cover, which is connected to the upper side of the kit body to close at least a portion of the plurality of chambers, and the cover includes a first limiting portion; The storage container includes a second limiting part. At the first position of the storage container, the second limiting part cooperates with the first limiting part to restrict the movement of the storage container in the vertical direction.

[0032] In some embodiments of the nucleic acid test kit, the first limiting part includes elastic claws; The second limiting part includes a limiting ring, which is disposed on the outer periphery of the storage container. At the first position of the storage container, the elastic claw is engaged with the bottom of the limiting ring, and at the second position of the storage container, the elastic claw is located above the storage container.

[0033] In some embodiments of the nucleic acid test kit, a third limiting part is provided in the reagent chamber corresponding to the storage container. In the second position, the third limiting part restricts the storage container from moving downward.

[0034] In some embodiments of the nucleic acid test kit, an ultrasonic mating part for cooperating with an ultrasonic device is provided below the extraction cavity, and the bottom surface of the ultrasonic mating part includes a plane or a downwardly convex arcuate surface.

[0035] A second aspect of this disclosure provides a nucleic acid testing system, including the nucleic acid testing cartridge of the above embodiments.

[0036] A third aspect of this disclosure provides a sample processing method for a nucleic acid detection cartridge based on the above embodiments, comprising: Step S10: Add the sample to be tested into the sample chamber; Step S20: The sample to be tested is lysed into the desired lysate within the extraction chamber; Step S30: The cleaning solution in the cleaning chamber is transported to the extraction chamber to clean the lysate, and the purification medium in the extraction chamber is used to adsorb nucleic acid to form a lysate suspension and the waste liquid after the nucleic acid adsorption is completed is discharged. Step S40: The eluent in the elution chamber is transferred to the extraction chamber to elute the adsorbate adsorbed by the purification medium to form an elution product; Step S50: The elution product is transferred to one of the lyophilized bulb chambers to rehydrate the lyophilized bulb therein, forming a first amplification reaction solution; and Step S60: The first amplification reaction solution is delivered to the amplification chamber, and a PCR amplification reaction is performed in the amplification chamber to form the first amplification product.

[0037] In some embodiments of the sample processing method, the plurality of reagent chambers include two lyophilized bulb chambers, wherein, after step S60, the sample processing method further includes: Step S70: The first amplification product formed in step S60 is delivered to the elution chamber, and the first amplification product is diluted with the elution solution in the elution chamber. Step S80: The diluted first amplification product is delivered to the other of the two lyophilized bulb chambers to re-dissolve the lyophilized bulb therein to form a second amplification reaction solution; Step S90: The second amplification reaction solution is delivered to the amplification chamber, and a PCR amplification reaction is performed in the amplification chamber to form a second amplification product.

[0038] In some embodiments of the sample processing method, Step S10 includes: adding the sample or a sample mixture of the sample and the preservative into the sample cavity as the sample to be tested; Step S20 includes: transferring the sample to be tested in the sample chamber and the lysis buffer in the lysis chamber to the extraction chamber to lyse the sample to be tested, using the purification medium in the extraction chamber to adsorb nucleic acid and discharging the waste liquid after nucleic acid adsorption.

[0039] In some embodiments of the sample processing method, Step S10 includes: processing the sample with a lysis buffer other than the nucleic acid detection cartridge to form the sample to be tested, and adding the sample to be tested into the sample chamber; Step S20 includes: transferring the sample to be tested from the sample chamber to the extraction chamber in multiple transfers, performing nucleic acid adsorption after each transfer and discharging the waste liquid after nucleic acid adsorption, until all samples to be tested are processed.

[0040] The nucleic acid detection cartridge provided in this embodiment includes a cartridge body, a rotary valve, and a drive unit. The cartridge body has a compartment section, which is divided into a transfer chamber, a sample chamber, multiple reagent chambers, and an extraction chamber by multiple partition walls. The multiple reagent chambers are used to pre-store lysis buffer, lyophilized bulbs, washing buffer, or elution buffer, etc. The rotary valve is rotatably disposed on the lower side of the cartridge body and has a first flow channel inside, selectively connecting the transfer chamber to any of the receiving chambers. The transfer chamber has a drive unit for driving fluid flow between the transfer chamber and the receiving chamber connected via the rotary valve. This nucleic acid detection cartridge integrates multiple nucleic acid detection steps, such as sample lysis, adsorption, washing, elution, and amplification reaction solution preparation, within the cartridge body. This not only results in a compact structure but also improves the operational continuity between the extraction steps, thus enhancing nucleic acid extraction efficiency.

[0041] The nucleic acid detection system disclosed herein includes the above embodiments, and thus has the advantages of the above embodiments of this disclosure.

[0042] The nucleic acid detection method provided in this disclosure is based on the nucleic acid detection system of the above embodiments, which can improve the operational coherence between multiple extraction steps, including sample lysis, adsorption, washing and elution, thereby improving the efficiency of nucleic acid extraction.

[0043] Other features and advantages of this disclosure will become clear from the following detailed description of exemplary embodiments with reference to the accompanying drawings. Attached Figure Description

[0044] The accompanying drawings, which are included to provide a further understanding of this disclosure and form part of this application, illustrate exemplary embodiments of this disclosure and are used to explain this disclosure, but do not constitute an undue limitation of this disclosure. In the drawings: Figure 1 This is a schematic diagram of the structure of a nucleic acid test cartridge according to some embodiments of this disclosure.

[0045] Figure 2 for Figure 1 The image shows an exploded view of the nucleic acid test kit.

[0046] Figure 3 for Figure 1 The front view of the main body of the nucleic acid test kit shown.

[0047] Figure 4 for Figure 1 The image shows a side cross-sectional view of the nucleic acid test kit with the lid open.

[0048] Figure 5 for Figure 1 The image shows a partial cross-sectional view of the nucleic acid test cartridge, which reveals the cartridge slot.

[0049] Figure 6 for Figure 1 The nucleic acid test cartridge shown is a cross-sectional view of the first plane, which shows the connected liquid flow channel and the first flow channel.

[0050] Figure 7 for Figure 1 The nucleic acid test cartridge shown is a cross-sectional view in the second plane, which shows the connected liquid flow channel and the second flow channel.

[0051] Figure 8 for Figure 1 The image shows a top view of the main body of the nucleic acid test kit.

[0052] Figure 9 for Figure 1 The image shows a bottom view of the main body of the nucleic acid test kit.

[0053] Figure 10 for Figure 1 The diagram shows a partial cross-sectional view of the main body of the nucleic acid test kit.

[0054] Figure 11 for Figure 1 The image shows a bottom view of the nucleic acid test kit with the lid open.

[0055] Figure 12 for Figure 1 The diagram shows a partial cross-sectional view of the lower cover of the nucleic acid test card box.

[0056] Figure 13 for Figure 1 The diagram shows a top view of the rotary valve of the nucleic acid test cartridge.

[0057] Figure 14 for Figure 13 The diagram shows a cross-sectional view of the rotary valve of the nucleic acid test cartridge in a first direction, which reveals the first flow channel.

[0058] Figure 15 for Figure 13 The cross-sectional view of the rotary valve of the nucleic acid test cartridge shown is in the second direction, which reveals the second flow channel.

[0059] Figure 16 for Figure 1 The image shows a side view of the amplification chip in the nucleic acid test cartridge.

[0060] Figure 17This is a schematic diagram of the structure of a nucleic acid test cartridge according to other embodiments of this disclosure.

[0061] Figure 18 for Figure 17 The diagram shows a partial cross-sectional view of the nucleic acid detection cartridge in the embodiment shown.

[0062] Figure 19 for Figure 18 The diagram shows a cross-sectional view of the nucleic acid test kit.

[0063] Figure 20 This is a schematic diagram of the main body of a nucleic acid testing cartridge according to other embodiments of this disclosure.

[0064] Figure 21 This is a schematic diagram of the main body of the nucleic acid detection cartridge and a portion of the ultrasonic device of the nucleic acid detection system according to some embodiments of this disclosure.

[0065] Explanation of reference numerals in the attached figures: 1. Base; 10. Base body; 11. Side limiting structure; 111. First inclined surface; 2. Cartridge body; 20. Extraction chamber; 21. Lysis chamber; 22. First washing chamber; 23. Second washing chamber; 24. Third washing chamber; 25. Elution chamber; 27. First lyophilization bulb chamber; 28. Second lyophilization bulb chamber; 200. Chamber body; 201. Transfer chamber; 202. Extraction chamber connection hole; 203. Lysis chamber connection hole; 204. First washing solution chamber connection hole; 205. Second washing solution chamber connection hole; 206. Third washing solution chamber connection hole; 207. Elution chamber connection hole; 208. First amplification chamber connection hole; 209. Second amplification chamber connection hole; 210. Gas chamber connection hole; 211. First lyophilization bulb chamber connection hole; 212. Second lyophilization bulb chamber connection hole; 213. Sample chamber connection hole; 214. Transfer chamber connection hole; 215. Waste liquid chamber connection hole. 216. Inlet hole; 217. Vent hole; 218. Partition wall; 219. Sub-partition wall; 220. Mounting part; 221. Mounting section; 222. Connecting section; 230. Rotary valve mating part; 241. Spiked base; 242. Spike tooth; 250. Ultrasonic mating part; 261. Second air chamber; 262. First air chamber; 271. First welding part; 280. First snap-fit ​​part; 291. Sample chamber; 292. Waste liquid chamber; 2910. Filter element; 2921. Waste liquid sub-chamber; 2922. Drying sub-chamber; 2a. Drainage hole; 2b. First vent; 2c. Second vent; 2d. Third vent; 20a. Arc-shaped surface; 27a. Third limiting part; 200a. Liquid flow channel; 200b. Gas flow channel; 220a. Slot; 3. Cover body; 31. Upper cover body; 311. Sealing part; 312. Upper cover body opening; 32. Lower cover body; 321. Second welding part; 322. Lower cover body opening; 34. Elastic claw; 3a. Cover body sealing film; 30a. Sealing film opening; 4. Amplification chip; 40. Amplification chamber; 401. Liquid passage; 4011. Large diameter section; 4012. Small diameter section; 4013. Transition section; 42. Gas passage; 411. Gas passage port fitting; 412. Liquid passage port fitting; 43. Connector; 431. Second inclined surface; 432. Connector body; 433. Side wing; 4a. Amplification chamber sealing film; 5. Rotary valve; 50. First flow channel; 51. Second flow channel; 50a. First flow channel port; 51a. Second flow channel port; 6. Drive unit; 7. Storage container; 71. Limiting ring; 73. Sealing part; 700. Ultrasonic equipment; A. Up and down direction; B. Front and back direction. Detailed Implementation

[0066] The technical solutions of the embodiments of this disclosure will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some embodiments of this disclosure, and not all embodiments. The following description of at least one exemplary embodiment is merely illustrative and is in no way intended to limit this disclosure or its application or use. All other embodiments obtained by those skilled in the art based on the embodiments of this disclosure without creative effort are within the scope of protection of this disclosure.

[0067] Unless otherwise specifically stated, the relative arrangement, numerical expressions, and values ​​of the components and steps set forth in these embodiments do not limit the scope of this disclosure. It should also be understood that, for ease of description, the dimensions of the various parts shown in the drawings are not drawn to actual scale. Techniques, methods, and devices known to those skilled in the art may not be discussed in detail, but where appropriate, such techniques, methods, and devices should be considered part of the specification. In all examples shown and discussed herein, any specific values ​​should be interpreted as merely exemplary and not as limitations. Therefore, other examples of exemplary embodiments may have different values. It should be noted that similar reference numerals and letters in the following drawings denote similar items; therefore, once an item is defined in one drawing, it need not be further discussed in subsequent drawings.

[0068] In the description of this disclosure, it should be understood that the use of terms such as "first" and "second" to define components is merely for the purpose of distinguishing the corresponding components. Unless otherwise stated, the above terms have no special meaning and therefore should not be construed as limiting the scope of protection of this disclosure.

[0069] In the description of this disclosure, it should be understood that the orientation or positional relationship indicated by directional terms such as "front, back, up, down, left, right", "horizontal, vertical, vertical, horizontal" and "top, bottom" are generally based on the orientation or positional relationship shown in the accompanying drawings, and are only for the convenience of describing this disclosure and simplifying the description. Unless otherwise stated, these directional terms do not indicate or imply that the device or element referred to must have a specific orientation or be constructed and operated in a specific orientation, and therefore should not be construed as a limitation on the scope of protection of this disclosure; the directional terms "inner" and "outer" refer to the inner and outer contours relative to the outline of each component itself.

[0070] In the following description, "before" refers to Figure 4 and Figure 5 "Left" refers to the left side; "back" refers to the side opposite to "front," and "left" and "right" refer to the left and right directions formed when facing forward.

[0071] like Figures 1 to 17 As shown, this embodiment of the present disclosure provides a nucleic acid detection cartridge. The nucleic acid detection cartridge includes a cartridge body 2, a rotary valve 5, an amplification chip 4, and a drive unit 6. The cartridge body 2 includes a compartment 200. The compartment 200 includes an outer wall enclosing an internal space and a plurality of partition walls 218 located in the internal space and extending in a vertical direction A. The plurality of partition walls 218 are configured to divide the internal space of the compartment 200 into a plurality of chambers. The plurality of chambers include a transfer chamber 201 and a plurality of receiving chambers. The plurality of receiving chambers include a sample chamber 291, a plurality of reagent chambers, and an extraction chamber 20. The plurality of reagent chambers include a lysis chamber 21 pre-stored with lysis buffer, a washing chamber pre-stored with washing buffer, an elution chamber 25 pre-stored with elution buffer, and a lyophilized bulb chamber pre-stored with lyophilized bulbs. The amplification chip 4 is mounted on the cartridge body 2 and includes an amplification chamber 40. A rotary valve 5 is rotatably disposed on the lower side of the cartridge body 2 and includes a first flow channel 50, which is configured to selectively connect the transfer chamber 201 to any receiving chamber or amplification chamber 40. A drive unit 6 is located in the transfer chamber 201 and is configured to drive fluid to flow between the transfer chamber 201 and the receiving chamber connected through the rotary valve 5.

[0072] The system includes multiple reagent chambers pre-stored with lysis buffer, washing buffer, elution buffer, and lyophilized beads, with each reagent pre-stored in its corresponding chamber. The lysis buffer disrupts the structure of cells or viruses to release internal nucleic acids while inhibiting the activity of degrading enzymes to prevent damage to the target molecule. Lyophilized beads are solid microspheres made by pre-mixing and freeze-drying amplification reagents such as enzymes and primers. The lyophilized reagents can be rapidly reconstituted with a suitable solvent. The washing buffer removes impurities from the nucleic acids. The elution buffer provides a low-salt buffer environment, dissolving and releasing nucleic acids bound to purification media, such as magnetic beads.

[0073] For example, the amplification chip 4 may include one amplification chamber 40, or multiple amplification chambers 40 to support multiplex nucleic acid detection.

[0074] For example, lyophilized bulbs can be pre-stored in one reagent chamber or in two separate reagent chambers. As another example, cleaning solution can be pre-stored in at least two reagent chambers. The transfer chamber 201 and the volumes of the various containing chambers can be the same or different.

[0075] like Figure 2 As shown, the drive unit 6 is, for example, a piston, which is made of rubber. When using the nucleic acid test cartridge, after the nucleic acid test cartridge is placed in the mounting position of the nucleic acid test system, the piston can be driven to the first drive element of the nucleic acid test system, such as an electric push rod, so that the piston can be driven to reciprocate in the vertical direction A within the transfer chamber 201 by the first drive element. The rotary valve 5 can be driven to the second drive element of the nucleic acid test system, such as a rotary motor, so that the rotary valve 5 can be driven to rotate relative to the cartridge body 2 by the second drive element, so as to select the receiving cavity communicating with the transfer chamber 201.

[0076] The nucleic acid detection cartridge provided in this embodiment includes a cartridge body 2, a rotary valve 5, and a drive unit 6. The cartridge body 2 has a compartment 200, which is divided into multiple chambers by multiple partition walls 218. The multiple chambers include a transfer chamber 201 and multiple receiving chambers. The multiple receiving chambers include a sample chamber 291, multiple reagent chambers, and an extraction chamber 20. The multiple reagent chambers are used to pre-store lysis buffer, lyophilized bulbs, washing buffer, or elution buffer, etc. The rotary valve 5 is rotatably disposed on the lower side of the cartridge body 2 and has a first flow channel 50 inside, which can selectively connect the transfer chamber 201 to any of the receiving chambers. The transfer chamber 201 is provided with a drive unit 6, which is used to drive fluid to flow between the transfer chamber 201 and the receiving chamber connected via the rotary valve 5. This nucleic acid detection cartridge integrates multiple nucleic acid detection steps, including sample lysis, adsorption, washing, elution, and amplification reaction solution preparation, into the cartridge body 2. This not only results in a compact structure but also improves the operational continuity between each extraction step, thus enhancing nucleic acid extraction efficiency. Furthermore, the lyophilized pellets help avoid the dust and cross-contamination risks associated with traditional lyophilized powders during dispensing. The independent granular shape of the pellets also prevents them from adhering to the reagent chamber wall, facilitating thorough reconstitution. For example, lyophilized pellets containing nucleic acid templates can be used for reconstitution, directly forming the amplification reaction solution within the reagent chamber, thereby improving nucleic acid extraction efficiency.

[0077] like Figure 2 As shown, in some embodiments, the nucleic acid detection cartridge also includes an amplification chip 4, which is mounted on the cartridge body 2 and includes an amplification cavity 40. The first flow path 50 of the rotary valve 5 is configured to selectively connect the intermediate transfer chamber 201 to the amplification chamber 40.

[0078] like Figure 4 and Figure 6 As shown, in some embodiments, the chamber 200 further includes a liquid flow channel 200a, the amplification chamber 40 is connected to the liquid flow channel 200a, and the first flow channel 50 of the rotary valve 5 is configured to selectively connect the intermediate transfer chamber 201 and the liquid flow channel 200a.

[0079] like Figure 1 , Figure 2 , Figure 4 and Figure 5 As shown, in some embodiments, the nucleic acid test kit further includes a base 1, which is connected to the lower side of the kit body 2. The kit body 2 also includes a rotary valve mating part 230, which is made of elastic material and integrally formed on the lower side of the compartment part 200, forming a rotary valve cavity between it and the base 1. The rotary valve 5 is disposed in the rotary valve cavity and is sealed to the rotary valve mating part 230.

[0080] The chamber body 200 can be made of a plastic material, such as polypropylene (PP).

[0081] The base 1 and the compartment 200 are detachably connected. For example... Figure 2 As shown, the chamber body 200 may include a first snap-fit ​​portion 280, located at the bottom of the chamber body 200. The first snap-fit ​​portion 280 may be, for example, a claw, a buckle, a flange, or a protrusion. The base 1 may include a second snap-fit ​​portion, configured to engage with the first snap-fit ​​portion. The second snap-fit ​​portion may be, for example, a groove or a hole. The first snap-fit ​​portion 280 and the second snap-fit ​​portion facilitate rapid assembly of the nucleic acid detection cartridge while ensuring the connection strength and accuracy between the base 1 and the chamber body 200 to maintain the seal between the rotary valve 5 and the rotary valve mating portion 230.

[0082] The rotary valve 5 is generally made of a rigid material (such as polycarbonate), while the rotary valve mating part 230 is made of an elastic material and integrally formed on the lower side of the housing part 200. On the one hand, this arrangement facilitates the formation of a tight fit between the contact surfaces of the rotary valve 5 and the rotary valve mating part 230 by utilizing the deformation capability of the elastic material, thereby achieving a reliable sealing and leak-proof effect. On the other hand, the buffering effect of the elastic material can absorb vibration and reduce wear or structural deformation caused by friction during the rotation of the rotary valve 5, thereby improving the service life and rotational stability of the rotary valve 5.

[0083] In addition, the base 1 can provide stable support for the rotary valve 5, which is also conducive to improving the rotational stability of the rotary valve 5.

[0084] like Figure 3 As shown, in some embodiments, the cartridge body 2 further includes a mounting portion 220, which is made of an elastic material and integrally formed on the front sidewall of the compartment portion 200. The mounting portion 220 is provided with a through hole for connecting the first flow channel 50 and the amplification cavity 40 of the amplification chip 4. The mounting portion 220 is used for sealing and engaging with the amplification chip 4.

[0085] like Figure 3 As shown, the mounting section 220 may include a mounting section 221 and a connecting section 222.

[0086] At least a portion of the edge of the mounting section 221 is oblong, and it is provided with two through holes, namely a liquid inlet 216 and a vent 217. The liquid inlet 216 and the vent 217 extend in the front-rear direction B and are spaced apart from each other in the vertical direction A. The liquid inlet 216 is used to connect the amplification chamber 40 and the first flow channel 50 of the rotary valve 5 (through the liquid flow channel 200a on the chamber body 200), and the vent 217 is used to connect the amplification chamber 40 and the second gas chamber 261 of the chamber body 200 (through the gas flow channel 200b on the chamber body 200).

[0087] The connecting section 222 can be elongated, with its first end connected to the bottom of the mounting section 221 and its second end connected to the rotary valve mating part 230. The connecting section 222 allows the mounting part 220 to be integrally formed with the rotary valve mating part 230, reducing processing steps and improving production efficiency.

[0088] like Figure 16 As shown, the amplification chip 4 includes a liquid channel port 412 and an air channel port 411. The liquid channel port 412 is inserted into the liquid inlet 216 and sealed to the cartridge body 2. Liquid in the transfer chamber 201, such as the amplification reaction solution, can be injected into the amplification chamber 40 through the first flow channel 50 and the liquid channel port 412 inserted into the liquid inlet 216. The air channel port 411 is inserted into the air outlet 217 and sealed to the cartridge body 2, so that when liquid is injected into the amplification chamber 40, the air in the amplification chamber 40 is discharged through the air channel port 411 inserted into the air outlet 217, so that the liquid can be smoothly injected into the amplification chamber 40.

[0089] The connection between the amplification chip 4 and the mounting part 220 is made of a rigid material (such as PP), while the mounting part 220 is made of an elastic material and is integrally formed on the front side wall of the compartment part 200. This is beneficial to utilize the deformation ability of the elastic material to form a tight fit between the adjacent surfaces of the two materials, thereby improving the sealing between the cartridge body 2 and the amplification chip 4.

[0090] In some embodiments, the valve mating portion 230 and / or mounting portion 220 are made of thermoplastic elastomer (TPE), thermoplastic polyurethane elastomer rubber (TPU), or liquid silicone material; and / or

[0091] The housing section 200 is made of PP or cyclic olefin copolymer (COC) or polycarbonate (PC); and / or

[0092] The valve mating part 230 and the housing part 200 are injection molded in two colors, and / or the mounting part 220 and the housing part 200 are injection molded in two colors; and / or

[0093] The surface of the rotary valve cavity formed by the rotary valve mating part 230 is in sync with the shape of the rotary valve 5.

[0094] The material used for the compartment body 200 has high hardness and chemical resistance, which helps to provide stable structural support. In addition, the selection of this material helps to improve the adhesion between the compartment body 200 and the valve mating part 230 and the mounting part 220, and is suitable for one-time molding using a two-color injection molding process, which helps to eliminate other bonding steps (such as adhesives, welding, etc.), thereby improving assembly efficiency and avoiding the risk of contamination.

[0095] In addition, the surface of the rotary valve mating part 230 that forms the rotary valve cavity matches the shape of the rotary valve 5, which also helps to improve the sealing performance.

[0096] like Figure 5 As shown, in some embodiments, the base 1 includes a side limiting structure 11 located on the front side of the mounting portion 220, and a slot 220a is formed between the side limiting structure 11 and the mounting portion 220. The slot 220a is configured to accommodate a connector 43 located on the rear side of the amplification chip 4, and the mounting portion 220 is configured to seal with the connector 43.

[0097] During assembly, the amplification chip 4 is first inserted into the mounting part 220, the liquid channel port 412 is connected to the liquid inlet 216, and the air channel port 411 is connected to the air outlet 217. Then, the base 1 and the cartridge body 2 are assembled to press the connector 43 of the amplification chip 4 into the slot 220a between the side limiting structure 11 and the mounting part 220, so that the amplification chip 4 is firmly and sealed on the cartridge body 2.

[0098] The connector 43 is inserted into the card slot 220a and has an interference fit with the mounting part 220. This helps to improve the connection strength between the amplification chip 4 and the base 1 and the card box body 2, and also helps to improve the sealing strength and prevent liquid or air leakage.

[0099] like Figure 5 As shown, the rear surface of the side limiting structure 11 includes a first inclined surface 111. The first inclined surface 111 forms an acute angle with the vertical direction A and gradually slopes backward from top to bottom. The first inclined surface 111 is configured to match the shape of the second inclined surface 431 provided on the front surface of the connector 43.

[0100] Further as Figure 5 As shown, the base 1 may also include a base body 10. The side limiting structure 11 includes a first limiting segment and a second limiting segment. The first limiting segment extends along the front-rear direction B, and its rear end is connected to the front side of the base body 10. The second limiting segment extends along the vertical direction A, and its first end is connected to the front end of the first limiting segment, which is a free end. The rear wall of the second limiting segment forms a first inclined surface 111.

[0101] Further as Figure 2 and Figure 5 As shown, the connector 43 includes a connector body 432 and two side wings 433. The two side wings 433 are connected to the left and right sides of the connector body 432, and the rear wall of the side wings 433 forms a second inclined surface 431.

[0102] The first inclined surface 111 and the second inclined surface 431 are designed to fit together, which helps to guide the connector 43 to slide smoothly into the slot 220a during assembly. After the connector is inserted, the squeezing force between the first inclined surface 111 and the second inclined surface 431 helps to make the connector 43 fit more tightly with the base 1, thereby helping to prevent the structure from loosening.

[0103] like Figure 8 and Figure 9 As shown, in some embodiments, the cartridge body 2 includes a plurality of drainage holes 2a, which are correspondingly arranged with a plurality of receiving cavities and are capable of communicating with the first flow channel 50. The drainage holes 2a include a narrowing section whose diameter gradually decreases from top to bottom. The bottom wall of at least one receiving cavity includes an inclined bottom surface that gradually slopes downwards from the bottom end of the partition wall 218 surrounding the receiving cavity to the drainage hole 2a. The narrowing section is tapered.

[0104] like Figure 8 and Figure 9 As shown, in some embodiments, the drainage hole 2a includes a first section located on the bottom wall of the chamber portion 200 and a second section located within the rotary valve mating portion 230. The diameter of the orifice of the second section away from the first section can be equal to the diameter of the first flow channel 50. The diameter of the first flow channel 50 can range from 0.5mm to 1.0mm, for example, 0.8mm. Appropriately setting the diameter range of the first flow channel 50 helps to ensure smooth liquid flow within a limited space.

[0105] The drainage hole 2a has a narrowed section, and the bottom wall of at least one receiving cavity gradually slopes downwards from the area away from the drainage hole 2a. This design serves to guide the flow of liquid, facilitating rapid inflow of liquid into the drainage hole 2a and improving the liquid reagent transfer rate between the receiving cavities. Furthermore, this design helps reduce residual liquid reagent in each receiving cavity, preventing insufficient sample dosage after extraction from affecting nucleic acid detection results.

[0106] like Figure 8 As shown, in some embodiments, the multiple reagent chambers include multiple cleaning chambers, each pre-stored with a different cleaning solution; and / or

[0107] Multiple reagent chambers include multiple lyophilized bulb chambers.

[0108] Different washing solutions can remove proteins, salt ions and other impurities bound to the purification medium in steps, thereby improving the purity of nucleic acids.

[0109] For example, the multiple reagent chambers may include three cleaning chambers (first cleaning chamber 22, second cleaning chamber 23 and third cleaning chamber 24) and two lyophilized bulb chambers (first lyophilized bulb chamber 27 and second lyophilized bulb chamber 28).

[0110] Setting up multiple washing chambers allows for the pre-storage of various washing solutions. By repeatedly using washing solutions with different compositions (e.g., first washing away impurities such as proteins and polysaccharides with a high-salt, low-pH solution, then desalting with an alcohol-containing solution, and finally eluting with a low-salt, high-pH solution to obtain purified nucleic acid templates), it is beneficial to reduce interference from impurities, improve the purity of the extracted nucleic acid, and thus improve the accuracy of nucleic acid detection results.

[0111] Nested PCR requires multiple amplifications. Setting up multiple lyophilized bulb cavities allows lyophilized bulbs to be pre-stored in different cavities of the nucleic acid detection cartridge simultaneously, making the nucleic acid detection cartridge suitable for nested PCR.

[0112] like Figure 17 As shown, in some embodiments, multiple reagent chambers may include a lyophilized bulb chamber, and a storage container 7 is provided inside the lyophilized bulb chamber. The storage container 7 is used to store the lyophilized bulb, which helps to prevent the lyophilized bulb from getting damp and also helps to pre-package the lyophilized bulb in the nucleic acid detection card box, eliminating the need for separate storage and transportation of the lyophilized bulb.

[0113] In some embodiments, such as Figure 8 As shown, the multiple receiving chambers may include a sample chamber 291, a lysis chamber 21, a first washing chamber 22, a second washing chamber 23, a third washing chamber 24, an elution chamber 25, an extraction chamber 20, a waste liquid chamber 292, a first lyophilized bulb chamber 27, and a second lyophilized bulb chamber 28. Multiple drainage holes 2a may be distributed circumferentially, and the included angle between two adjacent drainage holes 2a circumferentially may be an acute angle. For example, the multiple drainage holes 2a may include an extraction chamber connection hole 202, a lysis chamber connection hole 203, a first washing liquid chamber connection hole 204, a second washing liquid chamber connection hole 205, a third washing liquid chamber connection hole 206, an elution chamber connection hole 207, a first lyophilized bulb chamber connection hole 211, a second lyophilized bulb chamber connection hole 212, a sample chamber connection hole 213, and a waste liquid chamber connection hole 215. The extraction chamber connection hole 202 is located at the bottom of the extraction chamber 20. The lysis chamber connection hole 203 is located at the bottom of the lysis chamber 21. The first washing chamber connection hole 204, the second washing chamber connection hole 205, and the third washing chamber connection hole 206 are located at the bottom of the three washing chambers, respectively. The elution chamber connection hole 207 is located at the bottom of the elution chamber 25. The first freeze-drying bulb connection hole 211 and the second freeze-drying bulb connection hole 212 are located at the bottom of the two freeze-drying bulb chambers, respectively. The sample chamber connection hole 213 is located at the bottom of the sample chamber 291. The waste liquid chamber connection hole 215 is located at the bottom of the waste liquid chamber 292.

[0114] In addition, such as Figure 9 As shown, the bottom of the cartridge body 2 is also provided with a first amplification chamber connection hole 208 and a second amplification chamber connection hole 209. The first amplification chamber connection hole 208 and the second amplification chamber connection hole 209 are respectively the ports of the liquid flow channel 200a and the gas flow channel 200b that are connected to the first flow channel 50 and the second flow channel 51 of the rotary valve 5.

[0115] For example, the included angle between the extraction chamber connection port 202 and the lysis chamber connection port 203 can be in the range of 25° to 30°, for example, 28°. The included angle between the lysis chamber connection port 203 and the first washing chamber connection port 204 can be in the range of 25° to 30°, for example, 28°. The included angle between the first washing chamber connection port 204 and the second washing chamber connection port 205 can be in the range of 25° to 30°, for example, 30°. The included angle between the second washing chamber connection port 205 and the third washing chamber connection port 206 can be in the range of 30° to 35°, for example, 34°. The included angle between the third washing chamber connection port 206 and the elution chamber connection port 207 can be in the range of 30° to 35°, for example, 33°. The included angle between the elution chamber connection port 207 and the first amplification chamber connection port 208 can be in the range of 15° to 20°, for example, 20°. The included angle between the first amplification chamber connection port 208 and the second amplification chamber connection port 209 can range from 10° to 15°, for example, 14°. The included angle between the second amplification chamber connection port 209 and the first lyophilized bulb connection port 211 can range from 20° to 25°, for example, 23°. The included angle between the first lyophilized bulb connection port 211 and the second lyophilized bulb connection port 212 can range from 35° to 40°, for example, 38°. The included angle between the second lyophilized bulb connection port 212 and the sample chamber connection port 213 can range from 40° to 45°, for example, 45°. The included angle between the sample chamber connection port 213 and the waste liquid chamber connection port 215 can range from 40° to 45°, for example, 41°. The included angle between the waste liquid chamber connection port 215 and the extraction chamber connection port 202 can range from 25° to 30°, for example, 26°.

[0116] like Figure 8 As shown, in some embodiments, the plurality of chambers further includes a first air chamber 262, which is separated from the sample chamber 291 by a partition wall 218; A first air passage 2b is opened at the top of the partition wall 218 that separates the first air chamber 262 and the sample chamber 291 to connect the sample chamber 291 and the first air chamber 262.

[0117] The first air chamber 262 can be formed between the multiple freeze-drying bulb cavities and the outer wall of the chamber body 200. The sample chamber 291 is located behind the first air chamber 262 and the multiple freeze-drying bulb cavities. This layout is conducive to compact space.

[0118] The first air chamber 262 and the first air outlet 2b can balance the air pressure in the sample chamber 291, which helps to ensure the smooth flow of liquid reagents between the sample chamber 291 and the transfer chamber 201.

[0119] like Figure 8 As shown, in some embodiments, the plurality of receiving cavities also include a waste liquid cavity 292, which is separated from the extraction cavity 20 by a partition wall 218; A second vent 2c is provided at the top of the partition wall 218 that separates the waste liquid chamber 292 and the extraction chamber 20. The second vent 2c connects the waste liquid chamber 292 and the extraction chamber 20.

[0120] Setting up a second vent 2c can balance the air pressure in the waste liquid chamber 292 and the extraction chamber 20, which helps to maintain the stability of the air pressure in the extraction chamber 20.

[0121] like Figure 8 , Figures 13 to 15 As shown, in some embodiments, the multiple chambers further include a second air chamber 261, which is configured to keep the air pressure value in the amplification chamber 40 within a preset air pressure range. The rotary valve 5 also includes a second flow channel 51, which is configured to connect the second gas chamber 261 and the amplification chamber 40 when the first flow channel 50 is in the state of connecting the rotary chamber 201 and the amplification chamber 40.

[0122] The preset air pressure range is 40° so that the liquid can smoothly enter and exit the amplification chamber.

[0123] like Figure 8 As shown, the second air chamber 261 is located on the front side of the cartridge body 2 and can be positioned between the first freeze-drying bulb chamber 27 and the elution chamber 25 in the left-right direction. This layout is conducive to the compact structure and rapid fluid transfer between the chamber 200 and the amplification chamber 40.

[0124] like Figure 7 As shown, in some embodiments, the chamber 200 further includes a gas flow channel 200b, the amplification chamber 40 is connected to the gas flow channel 200b, and the second flow channel 51 is configured to connect the amplification chamber 40 and the second gas chamber 261 through the gas flow channel 200b while the first flow channel 50 is connected to the transfer chamber 201 and the amplification chamber 40.

[0125] Further as Figures 13 to 15 As shown, the second flow channel 51 is U-shaped and includes two second flow channel ports 51a. Further as... Figure 9As shown, the bottom of the second gas chamber 261 has a gas chamber connection hole 210. The gas chamber connection hole 210 and the second flow channel port 51a near the radial center of the rotary valve 5 are arranged opposite each other in the vertical direction, and are connected to each other when the rotary chamber 201 and the amplification chamber 40 are connected in the first flow channel 50. The distance between the gas chamber connection hole 210 and the rotation center of the rotary valve 5 is less than the distance between each drainage hole 2a and the rotation center of the rotary valve 5. The other second flow channel port 51a is connected to the amplification chamber 40 through the second amplification chamber connection hole 209, the gas flow channel 200b, and the gas passage hole 217, so that the gas pressure of the amplification chamber 40 can be balanced through the second gas chamber 261.

[0126] The second gas chamber 261 and the second flow channel 51 are designed to balance the gas pressure in the amplification chamber 40, ensuring that liquid can be smoothly injected into the amplification chamber 40 from the transfer chamber 201 or flow back from the amplification chamber 40 to the transfer chamber 201.

[0127] like Figure 8 As shown, in some embodiments, the waste liquid chamber 292 is further provided with a partition wall 219, which divides the waste liquid chamber 292 into a waste liquid sub-chamber 2921 and a drying sub-chamber 2922. The drying sub-chamber 2922 is connected to the extraction chamber 20 through the second air outlet 2c. The first flow channel 50 is configured to selectively connect the transfer chamber 201 with the waste liquid sub-chamber 2921. A desiccant is placed in the drying sub-chamber 2922.

[0128] The waste liquid chamber 292 is used to receive waste liquid generated during nucleic acid extraction (e.g., waste liquid after cleaning sample reagents with cleaning agent). The waste liquid chamber 292 is divided into a waste liquid sub-chamber 2921 and a drying sub-chamber 2922 by a partition wall 219. This facilitates the use of the waste liquid sub-chamber 2921 to store waste liquid, and the gas in the extraction chamber 20 can enter the drying sub-chamber 2922 through the second gas outlet 2c for drying with a desiccant.

[0129] like Figure 8 As shown, in some embodiments, a partition wall 219 is disposed on the partition wall 218 separating the waste liquid chamber 292 and the extraction chamber 20; and / or

[0130] A third vent 2d is opened at the top of the partition wall 219, and the third vent 2d connects the waste liquid sub-cavity 2921 and the drying sub-cavity 2922.

[0131] like Figure 8 and Figure 10 As shown, in some embodiments, the card holder body 2 includes a first welding part 271, which is disposed along the top of the outer wall of the compartment body 200 and / or the top of the partition wall 218. The nucleic acid test kit also includes a cover 3, which is connected to the top of the kit body 2 to close at least some of the multiple chambers. The cover 3 includes a second welding part 321, which is located at the bottom end of the cover 3 and is configured to be interlocked and welded together with the first welding part 271.

[0132] like Figure 10 As shown, the first welded part 271 is provided conformally at the top of the square outer wall of the compartment part 200 and the top of the partition wall 218.

[0133] In this design, one of the first welding part 271 and the second welding part 321 can be a welding wire, and the other is a groove that matches the shape of the welding wire, so that they can fit together.

[0134] The second welding part 321 and the first welding part 271 are fitted together, which helps to improve the sealing between the cover 3 and the card box body 2. This helps to prevent air leakage from affecting the transfer of liquid reagents between the various cavities, or to prevent leakage from mixing different reagents. It also helps to prevent impurities from entering the compartment 200 and contaminating the reagents.

[0135] In some embodiments, such as Figure 1 and Figure 2 As shown, the cover 3 includes an upper cover 31, a lower cover 32, and a cover sealing film 3a. The lower cover 32 is sealed to the top of the outer wall of the compartment 200 and the top of the multiple partition walls 218, and includes multiple lower cover openings 322 that communicate with some of the multiple chambers, including a sample chamber 291 and a transfer chamber 201.

[0136] The cover sealing film 3a is connected above the lower cover 32 and includes two sealing film openings 30a respectively opposite to the sample chamber 291 and the transfer chamber 201, used to seal the lower cover openings 322 of the plurality of lower cover openings 322 that correspond to the lower cover openings 322 other than the sample chamber 291 and the transfer chamber 201; and

[0137] The upper cover 31 is movably connected to the lower cover 32, and has a snap-fit ​​state covering the lower cover 32 and the cover sealing film 3a, and an open state exposing the lower cover 32 and the cover sealing film 3a. It includes a sealing part 311 and an upper cover opening 312. In the snap-fit ​​state, the sealing part 311 is opposite to the sample cavity 291 and is configured to block the lower cover opening 322 and the sealing film opening 30a corresponding to the sample cavity 291. The upper cover opening 312 is opposite to the transfer cavity 201 and the sealing film opening 30a corresponding to the transfer cavity 201.

[0138] like Figure 2 , Figure 11 and Figure 12As shown, the upper cover 31 and the lower cover 32 have the same shape, for example, square. The upper cover 31 and the lower cover 32 are hinged to form a flip-top structure. The lower cover 32 is connected to the top of the card holder body 2 to close the lower cover opening 322. A second welded part 321 is provided at the bottom of the lower cover 32.

[0139] like Figure 2 and Figure 11 As shown, some chambers also include multiple washing chambers and elution chambers.

[0140] For example, the cover sealing film 3a can close the lower cover opening 322 corresponding to the cleaning chamber and the rinsing chamber.

[0141] like Figure 16 As shown, in some embodiments, the amplification chip 4 includes a liquid passage 401 for connecting the first flow channel 50 and the amplification cavity 40 and a gas passage 42 for keeping the gas pressure value in the amplification cavity 40 within a preset gas pressure range. The liquid passage 401 includes a large-diameter section 4011 away from the amplification cavity 40, a small-diameter section 4012 closer to the amplification cavity 40 with a diameter smaller than that of the large-diameter section 4011, and a transition section 4013 connecting the large-diameter section 4011 and the small-diameter section 4012, with the diameter gradually decreasing from the large-diameter section 4011 to the small-diameter section 4012.

[0142] The large diameter section 4011 has a large flow area, which helps to buffer the liquid reagent when it enters the liquid passage 401.

[0143] In some embodiments, such as Figure 17 As shown, the nucleic acid test kit also includes at least one storage container 7, which is movably located in one of the reagent chambers along the vertical direction A so as to be able to be in a first position and a second position in the reagent chamber; The cartridge body 2 includes a spike, which is located at the bottom of the reagent chamber corresponding to the storage container 7. At the first position of the storage container 7, the spike and the storage container 7 are spaced apart in the vertical direction A. At the second position of the storage container 7, the spike pierces the storage container 7.

[0144] For example, such as Figure 18 and Figure 19 As shown, the spike portion includes a spike base 241 and spike teeth 242. The spike base 241 is connected to the bottom of the card holder body 2, and the spike teeth 242 extend in the vertical direction. The bottom of the spike teeth 242 is connected to the top of the spike base 241, and the spike teeth 242 have a pointed tip.

[0145] The storage container 7 may have a sealing film at its bottom to facilitate rapid puncture by the spikes. The sealing film may be applied to the entire bottom of the storage container 7, or it may be applied to a portion of the bottom. The sealing film may be made of a polymer composite aluminum film.

[0146] Pressure can be applied to the storage container 7 to move it from a first position to a second position, and the bottom of the storage container 7 can be punctured by a spike. After the bottom of the storage container 7 is punctured, the liquid reagent can flow into or out of the storage container 7 through the puncture point, so that the liquid reagent can be transferred between the storage container 7 and the transfer chamber 201.

[0147] Further as Figure 17 As shown, the storage container 7 stores freeze-dried pellets. The outer wall of the storage container 7 can be conformally arranged to the freeze-dried pellet cavity, such as in a cylindrical shape, or it can also be a cylindrical structure such as a prism or cuboid, to facilitate the adaptability of the storage container 7 to the installation space.

[0148] The top of the storage container 7 can be sealed with a PP composite aluminum film, which eliminates the need for a lid structure and improves the sealing performance of the storage container 7. Alternatively, the top of the storage container 7 can also be sealed with a lid 3. Alternatively, the storage container 7 can be formed as an inverted bottomed cylindrical structure with a sealed film at the opening.

[0149] Before the bottom of storage container 7 is punctured by the spike, storage container 7 is in a sealed state, thus preventing impurities from contaminating the reagents inside, which helps improve the purity of nucleic acid extraction and consequently improves the accuracy of test results. In addition, this setup saves the process of manually adding and preparing reagents, reducing human error and the risk of cross-contamination, and also facilitates automated testing.

[0150] like Figure 17 As shown, in some embodiments, the nucleic acid test cartridge further includes a sealing portion 73, which is configured to seal the gap between the storage container 7 and the spike portion.

[0151] The sealing part 73 can be located near the bottom of the spike, for example, the sealing part 73 can be sleeved on the outer periphery of the spike base 241.

[0152] For example, the sealing part 73 can be made of materials such as rubber or PP to form sealing structures such as sealing rings and sealing sleeves.

[0153] Liquid enters storage container 7 from the bottom, and the sealing part 73 ensures the seal between storage container 7 and the spike, thereby helping to prevent impurities from entering storage container 7.

[0154] like Figure 19 As shown, in some embodiments, the sealing portion 73 includes at least one sealing ring disposed at the bottom of the storage container 7 and arranged circumferentially around the spike portion.

[0155] The sealing ring has a simple structure and is easy to manufacture. In addition, the sealing ring has good centering, which helps to position the storage container 7 in the receiving cavity and prevents leakage caused by shaking of the storage container 7 during reagent transfer.

[0156] like Figure 17 As shown, in some embodiments, the nucleic acid test kit further includes a cover 3, which is connected to the upper side of the kit body 2 to close at least a portion of the multiple receiving cavities. The cover 3 includes a first limiting part. The storage container 7 includes a second limiting part. In the first position of the storage container 7, the second limiting part cooperates with the first limiting part to restrict the movement of the storage container 7 in the vertical direction A.

[0157] For example, the first limiting part and the second limiting part can be a flange and groove that cooperate with each other, a buckle and a slot, a claw and a slot, etc.

[0158] The first and second limiting parts improve the structural stability of the storage container 7 in the first position, preventing it from becoming loose and accidentally punctured by the spikes, thus exposing the lyophilized reagent inside to the external environment and causing it to absorb moisture from the air. This can lead to the lyophilized reagent losing its activity, clumping, or even deteriorating, making it impossible for the expected reconstitution reaction to occur during subsequent sample injection, ultimately resulting in invalid test results.

[0159] like Figure 17 As shown, in some embodiments, the first limiting portion includes an elastic claw 34; The second limiting part includes a limiting ring 71, which is disposed on the outer periphery of the storage container 7. In a first position of the storage container 7, an elastic claw 34 engages with the bottom of the limiting ring 71. In a second position of the storage container 7, the elastic claw 34 is positioned above the storage container 7. Further as... Figure 17 As shown, the elastic claw 34 extends along the vertical direction A, and the bottom of the elastic claw 34 bends inward to form a claw-shaped structure.

[0160] During the process of moving the storage container 7 from the first position to the second position, the elastic claw 34 is opened and moved above the storage container 7. This setting helps to limit the upward displacement of the storage container 7 in the event that the storage container 7 bounces off the bottom.

[0161] The elastic claw 34 and the limiting ring 71 have a small structure and can adapt to the narrow space inside the nucleic acid detection cartridge. The cooperation between the elastic claw 34 and the limiting ring 71 can achieve stable positioning of the storage container 7.

[0162] like Figure 19 As shown, in some embodiments, a third limiting part 27a is provided in the reagent chamber corresponding to the storage container 7. In the second position, the third limiting part 27a restricts the storage container 7 from moving downward.

[0163] For example, the third limiting part 27a is a stepped surface of the stepped inner wall of the reagent chamber where the storage container 7 is located. Providing the third limiting part 27a helps to limit the second position of the storage container 7, preventing the storage container 7 from moving further downward beyond the second position and being impacted.

[0164] like Figure 20 and Figure 21 As shown, in some embodiments, an ultrasonic mating part 250 for cooperating with an ultrasonic device 700 is provided below the extraction cavity 20, and the bottom surface of the ultrasonic mating part 250 includes a flat surface or a downwardly protruding arcuate surface 20a.

[0165] The ultrasonic mating part 250 is used to mate with the ultrasonic equipment 700. The bottom surface of the ultrasonic mating part 250 is flat, which facilitates manufacturing and also helps to compact the space along the vertical direction. In addition, the bottom surface of the ultrasonic mating part 250 is a downwardly convex arc-shaped surface 20a, which helps to form an energy-concentrating effect at the bottom of the extraction cavity 20, generating a stronger cavitation effect and micro-jet impact, thereby improving the ultrasonic mixing effect.

[0166] This disclosure also provides a nucleic acid detection system, including a nucleic acid detection cartridge according to this disclosure.

[0167] The nucleic acid detection system of this disclosure has the advantages of the nucleic acid detection cartridge of this disclosure.

[0168] In some embodiments, the nucleic acid detection system further includes a detection device configured to detect whether the amplification chip 4 has reached a preset position.

[0169] Setting up a detection device helps to promptly determine the position of the amplification chip 4, ensuring that the amplification cavity 40 reaches the preset position and avoiding misalignment of the amplification cavity 40 that could affect the detection results.

[0170] This disclosure also provides a sample processing method. The sample processing method includes: Step S10: Add the sample to be tested into the sample chamber 291; Step S20: The sample to be tested is lysed into the required lysate within the extraction chamber 20; Step S30: The cleaning solution in the cleaning chamber is transported to the extraction chamber 20 to clean the lysate. The purification medium in the extraction chamber 20 is used to adsorb nucleic acid to form a lysate suspension and the waste liquid after the nucleic acid adsorption is completed is discharged. Step S40: The eluent in the elution chamber 25 is transferred to the extraction chamber 20 to elute the adsorbate adsorbed by the purification medium to form the elution product. Step S50: The elution product is transferred to a lyophilized bulb cavity to rehydrate the lyophilized bulb therein, forming the first amplification reaction solution; and In step S60, the first amplification reaction solution is delivered to the amplification chamber 40, and a PCR amplification reaction is performed in the amplification chamber 40 to form the first amplification product.

[0171] like Figure 4 and Figure 5 As shown, liquid reagents can be transferred between the transfer chamber 201 and multiple receiving chambers via the drive unit 6. For example, the drive unit 6 includes a piston that moves upward to create a negative pressure in the transfer chamber 201, thereby drawing liquid from the reagent chamber or sample chamber 291 or extraction chamber 20, which are connected to the first flow channel 50, into the transfer chamber 201; or the piston moves downward to create a positive pressure in the transfer chamber 201, thereby pushing the liquid in the transfer chamber 201 into the reagent chamber or sample chamber 291 or extraction chamber 20, which are connected to the first flow channel 50, so that the liquid is transferred between the receiving chambers via the rotary valve 5 and the transfer chamber 201.

[0172] The sample processing method of this disclosure can improve the operational coherence between multiple nucleic acid detection steps, including sample lysis, adsorption, washing, elution, and preparation of amplification reaction solution, thereby improving the efficiency of nucleic acid detection.

[0173] In some embodiments, the plurality of reagent chambers include two lyophilized bulb chambers, wherein, after step S60, the sample processing method further includes: Step S70: The first amplification product formed in step S60 is delivered to the elution chamber 25, and the first amplification product is diluted with the elution buffer in the elution chamber 25. Step S80: The diluted first amplification product is transferred to the other lyophilized bulb chamber of the two lyophilized bulb chambers to re-dissolve the lyophilized bulbs therein to form the second amplification reaction solution; In step S90, the second amplification reaction solution is delivered to the amplification chamber 40, and a PCR amplification reaction is performed in the amplification chamber 40 to form the second amplification product.

[0174] Setting up two freeze-drying bulbs is beneficial for nested PCR.

[0175] In some embodiments, the multiple reagent chambers may also include three or more lyophilized bulb chambers to achieve more than three amplification reactions.

[0176] In some embodiments, step S10 includes adding a sample or a sample mixture of a sample and a preservative into the sample chamber 291 as the sample to be tested; Step S20 includes: transferring the sample to be tested in the sample chamber 291 and the lysis buffer in the lysis chamber 21 to the extraction chamber 20 to lyse the sample to be tested, using the purification medium in the extraction chamber 20 to adsorb nucleic acid and discharging the waste liquid after nucleic acid adsorption.

[0177] This sample processing method integrates the lysis step into the nucleic acid detection cartridge, which allows for direct extraction after the sample to be tested has been lysed, thereby improving the sample extraction speed.

[0178] In some embodiments, step S10 includes: processing the sample with a lysis buffer other than the nucleic acid detection cartridge to form a sample to be tested, and adding the sample to be tested into the sample chamber 291; Step S20 includes: transferring the sample to be tested from the sample chamber 291 to the extraction chamber 20 in multiple transfers, performing nucleic acid adsorption after each transfer and discharging the waste liquid after nucleic acid adsorption, until all the samples to be tested are processed to form the required lysate.

[0179] By treating the sample with lysis buffer outside the nucleic acid detection cartridge and transferring the sample mixture multiple times, this sample processing method enables the nucleic acid detection cartridge to extract large-volume samples. It also allows the number of cycles to be determined based on the sample size and the volume of sample that can be adsorbed in a single cycle, which helps to improve the detection sensitivity of large-volume samples.

[0180] The following combination Figures 1 to 17 The nucleic acid detection cartridge and sample processing method of the present disclosure will be described in detail.

[0181] like Figure 1 and Figure 2 As shown, the nucleic acid test kit includes a base 1, a kit body 2, a cover 3, an amplification chip 4, a rotary valve 5, and a drive unit 6.

[0182] The cartridge body 2 is mounted above the base 1, forming a rotary valve chamber between them. A rotary valve 5 is rotatably located within the rotary valve chamber and includes a first flow channel 50. The cartridge body 2 includes a compartment 200. The compartment 200 includes an outer wall enclosing an internal space and multiple partition walls 218 located within the internal space and extending in the vertical direction A. The multiple partition walls 218 are configured to divide the internal space into a transfer chamber 201 and multiple chambers. The multiple chambers include a first gas chamber 262, a transfer chamber 201, and multiple receiving chambers. The multiple receiving chambers include a pyrolysis chamber 21 pre-stored with pyrolysis solution, a first cleaning chamber 22 pre-stored with a first cleaning solution, a second cleaning chamber 23 pre-stored with a second cleaning solution, a third cleaning chamber 24 pre-stored with a third cleaning solution, an elution chamber 25 pre-stored with eluent, a first freeze-dried bulb chamber 27 and a second freeze-dried bulb chamber 28 pre-stored with freeze-dried bulbs, and a waste liquid chamber 292 for receiving waste liquid.

[0183] The multiple chambers also include a first air chamber 262, which is connected to the sample chamber 291 through a first air outlet 2b located at the top of the partition wall 218 separating the first air chamber 262 and the sample chamber 291, so as to balance the air pressure in the sample chamber 291.

[0184] The drive unit 6 is located within the transfer chamber 201 and is configured to drive fluid to flow between the transfer chamber 201 and a receiving chamber connected via the rotary valve 5. The first flow channel 50 is configured to selectively connect the transfer chamber 201 to either receiving chamber.

[0185] The amplification chip 4 is mounted on the cartridge body 2 and includes an amplification chamber 40. The first flow channel 50 of the rotary valve 5 can selectively connect the intermediate transfer chamber 201 to the amplification chamber 40. This nucleic acid detection cartridge integrates the steps of lysis, washing, elution, and reconstitution in the nucleic acid extraction process, which is beneficial to improving the efficiency of nucleic acid detection. In addition, this nucleic acid detection cartridge includes three washing chambers and two lyophilized bulb chambers, which is beneficial to multiple washing of liquid reagents, improving the purity of nucleic acid extraction, and allowing liquid reagents to be reconstituted multiple times for lyophilized bulbs, thereby facilitating nested PCR.

[0186] The extraction chamber 20 contains magnetic beads as a purification medium, which are used to adsorb nucleic acids during lysis and washing.

[0187] The card holder body 2 is also equipped with a desiccant for moisture-proof preservation of the magnetic beads. The desiccant is placed in the waste liquid chamber 292, which is connected to the extraction chamber 20 through a second air vent 2c located at the top of the partition wall 218 separating the waste liquid chamber 292 and the extraction chamber 20.

[0188] like Figure 8 As shown, the transfer cavity 201 is a cylindrical receiving cavity with a diameter of 4.6 mm.

[0189] like Figures 3 to 5 , Figure 9 As shown, the card holder body 2 also includes a rotary valve mating part 230 and a mounting part 220. The rotary valve mating part 230 is integrally formed on the lower side of the compartment part 200, forming a rotary valve cavity between it and the base 1. The second hole section of the drainage hole 2a is provided on the rotary valve mating part 230. The rotary valve mating part 230 has the same shape as the rotary valve 5, and the two are sealed together.

[0190] like Figures 3 to 5 , Figure 9 As shown, the mounting portion 220 is integrally formed on the front sidewall of the housing portion 200. The amplification chip 4 forms a sealed fit with the mounting portion 220.

[0191] The mounting section 220 may include a mounting segment 221 and a connecting segment 222. The mounting segment 221 has two through holes extending in the front-rear direction B, namely a liquid inlet 216 and a vent 217, with the liquid inlet 216 located at the bottom of the vent 217. The liquid inlet 216 connects the amplification chamber 40 and the first flow channel 50, and the vent 217 connects the amplification chamber 40 and the second gas chamber 261. The connecting segment 222 is elongated, with its first end connected to the bottom of the mounting segment 221 and its second end connected to the front side of the rotary valve mating part 230.

[0192] The nucleic acid test cartridge is molded using a two-color injection molding process. The valve mating part 230 and the mounting part 220 are integrally molded and made of TPE material. Except for the valve mating part 230, the other parts of the cartridge body 2 and the base 1 are made of PP material. The valve 5 is made of polycarbonate (PC) material, and the process holes formed during the molding of the valve 5 are sealed by a secondary injection molding process. PP and PC materials are harder than TPE materials, meaning there is a difference in elastic modulus between the soft and hard materials, which allows the adjacent surfaces of TPE material and other materials to form a tight fit, thereby improving the structural sealing performance.

[0193] like Figure 2 As shown, the top edge of the compartment 200 is roughly square, and the dimensions of the top edge of the compartment 200 are 37mm × 37.5mm. The height of the card holder body 2 is 63.5mm.

[0194] like Figure 8 and Figure 9 As shown, the card holder body 2 has 10 drainage holes 2a, each corresponding to one of the receiving cavities and capable of communicating with the first flow channel 50. The 10 drainage holes 2a are distributed on the same circumference. The drainage holes 2a include a narrowing section whose diameter gradually decreases from top to bottom. Furthermore, the bottom wall of each receiving cavity gradually slopes downward from the distance from the drainage holes 2a to the drainage holes 2a.

[0195] like Figure 8 As shown, a filter element 2910 extending in the vertical direction is provided in the sample cavity 291. The filter element 2910 is connected above the drainage hole 2a in the sample cavity 291, so as to filter the sample before the sample or sample mixture is extracted and the sample is waiting to be tested, thereby improving the purity of nucleic acid extraction.

[0196] like Figures 13 to 15 As shown, the rotary valve 5 includes a first flow channel 50 and a second flow channel 51. A transfer cavity connection hole 214 is provided at the bottom of the transfer cavity 201. The first flow channel 50 is configured to connect the transfer cavity connection hole 214 and any drain hole 2a, so as to connect the transfer cavity 201 and the receiving cavity where the drain hole 2a is located.

[0197] like Figure 15 As shown, the second flow channel 51 has two second flow channel ports 51a. The second flow channel port 51a near the rotation center of the rotary valve 5 is connected to the second gas chamber 261 through the gas chamber connection hole 210. The other second flow channel port 51a can be connected to the amplification chamber 40 through the gas passage hole 217. Thus, when liquid reagents are injected into the amplification chamber 40, the gas discharged from the amplification chamber 40 can be guided into the second gas chamber 261 to smoothly transfer the liquid.

[0198] like Figure 2 , Figure 4 and Figure 5 As shown, the drive unit 6 includes a piston located within the transfer chamber 201. The piston moves upward, creating a negative pressure within the transfer chamber 201, thereby drawing liquid from the receiving chamber connected to the first flow channel 50 into the transfer chamber 201. The piston moves downward, creating a positive pressure within the transfer chamber 201, thereby pushing the liquid from the transfer chamber 201 into the receiving chamber connected through the first flow channel 50.

[0199] like Figure 8 As shown, a first air vent 2b is provided at the top of the partition wall 218 separating the first air chamber 262 and the sample chamber 291 to connect the sample chamber 291 and the first air chamber 262. A second air vent 2c is provided at the top of the partition wall 218 separating the waste liquid chamber 292 and the extraction chamber 20, connecting the waste liquid chamber 292 and the extraction chamber 20. During nucleic acid extraction, the chamber 200 is sealed, and liquid transfer can cause an imbalance in internal air pressure. Therefore, the first air vent 2b and the second air vent 2c can compensate for the air pressure and improve the stability of the liquid transfer process.

[0200] like Figure 8 As shown, the waste liquid chamber 292 is also provided with a partition wall 219, which divides the waste liquid chamber 292 into a waste liquid sub-chamber 2921 and a drying sub-chamber 2922. The drying sub-chamber 2922 is connected to the extraction chamber 20 through a second air outlet 2c. The first flow channel 50 is configured to selectively connect the transfer chamber 201 with the waste liquid sub-chamber 2921. A desiccant is placed in the drying sub-chamber 2922. A third air outlet 2d is opened at the top of the partition wall 219, which connects the waste liquid sub-chamber 2921 and the drying sub-chamber 2922.

[0201] like Figure 1 and Figure 11 As shown, the cover 3 includes an upper cover 31, a lower cover 32, and a cover sealing film 3a. The upper cover 31 and the lower cover 32 are hinged together. The cover sealing film 3a is heat-bonded to the upper cover 31. The lower cover 32 has lower cover openings 322 corresponding to the three cleaning chambers, elution chamber 25, transfer chamber 201, sample chamber 291, and lysis chamber 21, respectively. The opening diameter corresponding to the sample chamber 291 is 7.9 mm, which can accommodate pharyngeal swabs with a diameter of 5.5 mm-5.8 mm, facilitating direct insertion of the pharyngeal swab. When the upper cover 31 is fastened to the lower cover 32, the cover sealing film 3a can close the lower cover openings 322 on the lower cover 32 except those corresponding to the sample chamber 291 and the transfer chamber 201.

[0202] like Figure 8 , Figures 10 to 12As shown, the lower cover 32 of the cover 3 is welded to the top of the card holder body 2, and the lower cover 32 and the card holder body 2 are sealed by ultrasonic bonding. The card holder body 2 includes a first welded portion 271, which is located at the top of the card holder body 2. The bottom of the lower cover 32 is provided with a second welded portion 321, which is a groove configured to accommodate the first welded portion 271.

[0203] like Figure 2 and Figure 16 As shown, the amplification chip 4 includes an amplification chip body, an amplification cavity 40, two amplification cavity sealing films 4a, and a connector 43. The amplification chip body extends along the front-to-back direction B, and the amplification cavity 40 is disposed within the amplification chip body. The two amplification cavity sealing films 4a are consistent in shape with the amplification chip body and cover the left and right sides of the amplification chip body to seal the amplification cavity 40. The amplification cavity 40 of the amplification chip 4 is formed using a double-sided film thermopressing / laser bonding sealing process.

[0204] like Figure 16 As shown, the connector 43 includes a connector body 432 and two side wings 433. The connector body 432 is connected to the rear side of the amplification chip body. The two side wings 433 are connected to the left and right sides of the connector body 432, and the rear wall of the side wings 433 forms a second inclined surface 431.

[0205] like Figure 5 As shown, the base 1 includes a base body 10 and a side limiting structure 11. The side limiting structure 11 is located on the front side of the mounting part 220, and a groove 220a is formed between the side limiting structure 11 and the mounting part 220. The connector 43 is inserted into the groove 220a and is interference-fitted with the mounting part 220.

[0206] like Figure 5 As shown, the side limiting structure 11 includes a first limiting segment and a second limiting segment. The first limiting segment extends along the front-rear direction B, and its rear end is connected to the front side of the base body 10. The second limiting segment extends along the vertical direction A, and its first end is connected to the front end of the first limiting segment, which is a free end. The rear wall of the second limiting segment forms a first inclined surface 111. The first inclined surface 111 and the second inclined surface 431 are shaped to fit together.

[0207] The connector 43 is made of PP material, which is harder than the material of the mounting part 220, so that a tight seal can be formed between the two.

[0208] like Figures 17-19 In the illustrated embodiment, with Figures 1 to 16The embodiment shown differs in that the nucleic acid test kit also includes a storage container 7, which pre-stores lyophilized bulbs. The compartment 200 has only one lyophilized bulb cavity, and the storage container 7 is movably installed within this cavity along the vertical direction A. The storage container 7 has a first position and a second position.

[0209] like Figure 17 As shown, the card holder body 2 includes a spike (not shown) located at the bottom of the freeze-drying bulb cavity. In a first position of the storage container 7, the spike is spaced apart from the storage container 7 in the vertical direction A. In a second position of the storage container 7, the spike pierces the storage container 7.

[0210] like Figure 18 and Figure 19 As shown, the spike portion includes a spike base 241 and spike teeth 242. The spike base 241 is connected to the bottom of the card holder body 2, and the spike teeth 242 extend in the vertical direction. The bottom of the spike teeth 242 is connected to the top of the spike base 241, and the spike teeth 242 have a pointed tip.

[0211] like Figure 17 As shown, the storage container 7 is a cylindrical container. The top of the storage container 7 is sealed with a PP composite aluminum film. The storage container 7 includes a sealing part 73, which is an O-ring, located at the bottom of the storage container 7 and fitted around the outer periphery of the spike base 241 to seal the gap between the storage container 7 and the spike base 241.

[0212] like Figure 17 As shown, the storage container 7 includes a second limiting part, which is a limiting ring 71 disposed on the outer periphery of the storage container 7. The lower cover 32 includes a first limiting part, which is an elastic claw 34 extending in the vertical direction A. In the first position of the storage container 7, the elastic claw 34 is engaged below the limiting ring 71. Before sample processing, the storage container 7 is held in the first position by the elastic claw 34. During sample processing, the storage container 7 is driven to move downward, and the sealing film at the bottom of the storage container 7 is punctured by the barbs 242 of the spikes.

[0213] like Figure 19 As shown, a third limiting part 27a is provided in the reagent chamber corresponding to the storage container 7. In the second position, the third limiting part 27a restricts the storage container 7 from moving downward.

[0214] like Figure 20 and Figure 21 In the illustrated embodiment, with Figures 1 to 19 The difference in the illustrated embodiment is that an ultrasonic mating part for cooperating with the ultrasonic device 700 is provided below the extraction cavity 20, and the bottom surface of the ultrasonic mating part includes a flat surface or a downwardly protruding arcuate surface 20a.

[0215] The sample processing methods of two embodiments of this disclosure will be described in detail below.

[0216] Example 1: The lysis buffer is pre-stored in the lysis chamber 21, the first cleaning solution is pre-stored in the first cleaning chamber 22, the second cleaning solution is pre-stored in the second cleaning chamber 23, the third cleaning solution is pre-stored in the third cleaning chamber 24, the eluent is pre-stored in the elution chamber 25, the magnetic beads are stored in the extraction chamber 20, and the lyophilized pellets are pre-stored in the first lyophilized pellet chamber 27 and the second lyophilized pellet chamber 28, respectively.

[0217] The sample to be tested is added to the sample chamber 291. The sample to be tested can be a liquid sample or a mixture of the sample and a preservative. Then, the rotary valve 5 is rotated to connect the first flow channel 50 with the sample chamber 291, and the piston is then controlled to move upward to draw the sample into the transfer chamber 201. Then, the rotary valve 5 is rotated to connect the first flow channel 50 with the extraction chamber 20, and the piston is then controlled to move downward to push the sample into the extraction chamber 20.

[0218] Subsequently, the lysis buffer was transferred to the extraction chamber 20 in the same manner as the sample transfer. The sample and lysis buffer in the extraction chamber 20 were ultrasonically mixed, and the mixed liquid reagent was adsorbed by magnetic beads. The resulting waste liquid was then discharged into the waste liquid chamber 292 to form the desired lysis product.

[0219] Subsequently, the first cleaning is performed: the first cleaning solution is transferred to the extraction chamber 20 in the same way as the sample transfer to form a lysate suspension, followed by magnetic bead adsorption and the resulting waste liquid is discharged to the waste liquid chamber 292.

[0220] Subsequently, a second cleaning solution was used for a second cleaning, followed by a third cleaning solution for a third cleaning. After each cleaning, magnetic beads were adsorbed and waste liquid was discharged.

[0221] Subsequently, the eluent was transferred to the extraction chamber 20 in the same manner as the sample transfer to form the elution product by eluting the adsorbate of the magnetic beads.

[0222] Subsequently, the elution product was transferred to the first lyophilized bulb cavity 27 in the same manner as the sample transfer. The lyophilized bulb in the first lyophilized bulb cavity 27 was reconstituted to form the first amplification reaction solution, and then the first amplification reaction solution was transferred to the transfer cavity 201.

[0223] Subsequently, the rotary valve 5 is rotated to connect the first flow channel 50 with the amplification chamber 40. The piston is moved to transfer the first amplification reaction solution in the transfer chamber 201 into the amplification chamber 40 to carry out the first stage of amplification reaction and obtain the first amplification product.

[0224] Subsequently, rotate the rotary valve 5 to connect the first flow channel 50 with the elution chamber 25, move the piston to transfer the first amplification product into the elution chamber 25, and use the elution buffer in the elution chamber 25 to dilute the first amplification product.

[0225] Subsequently, the amplification product is transferred from the elution chamber 25 to the second lyophilized bulb chamber 28, and the lyophilized bulb in the second lyophilized bulb chamber 28 is reconstituted to form the second amplification reaction solution. Subsequently, the second amplification reaction solution was transferred into the transfer chamber 201.

[0226] Subsequently, the second amplification reaction solution in the transfer chamber 201 is pushed into the amplification chamber 40 to carry out the second stage of amplification reaction to form the second amplification product.

[0227] Example 2: Unlike Example 1, the sample is first lysed outside the nucleic acid detection cartridge to obtain the sample to be tested, and then added to the sample chamber 291. The sample to be tested is then transferred to the extraction chamber 20 multiple times for magnetic bead adsorption and waste liquid removal, until all the sample to be tested has been processed to form the required lysate.

[0228] Finally, it should be noted that the above embodiments are only used to illustrate the technical solutions of this disclosure and not to limit them; although this disclosure has been described in detail with reference to preferred embodiments, those skilled in the art should understand that modifications can still be made to the specific implementation of this disclosure or equivalent substitutions can be made to some technical features, all of which should be covered within the scope of the technical solutions claimed in this disclosure.

Claims

1. A nucleic acid testing kit, characterized in that, include: The main body (2) of the card box includes a compartment (200), the compartment (200) includes an outer wall that encloses an internal space and a plurality of partition walls (218) located in the internal space and extending in the vertical direction (A). The plurality of partition walls (218) are configured to divide the internal space into a plurality of chambers. The plurality of chambers include a transfer chamber (201) and a plurality of receiving chambers. The plurality of receiving chambers include a sample chamber (291), a plurality of reagent chambers and an extraction chamber (20). The plurality of reagent chambers include a lysis chamber (21) for pre-stored lysis buffer, a washing chamber for pre-stored washing buffer, an elution chamber (25) for pre-stored elution buffer and a lyophilized bulb chamber for pre-stored lyophilized bulbs. An amplification chip (4) is mounted on the cartridge body (2) and includes an amplification cavity (40). A rotary valve (5) is rotatably disposed on the lower side of the cartridge body (2) and includes a first flow channel (50), which is configured to selectively connect the transfer chamber (201) to any of the receiving chambers or the amplification chamber (40). and The drive unit (6), located in the transfer chamber (201), is configured to drive fluid to flow between the transfer chamber (201) and the receiving chamber connected by the rotary valve (5).

2. The nucleic acid detection cartridge according to claim 1, characterized in that, The chamber (200) further includes a liquid flow channel (200a), the amplification chamber (40) is connected to the liquid flow channel (200a), and the first flow channel (50) of the rotary valve (5) is configured to selectively connect the intermediate transfer chamber (201) to the liquid flow channel (200a).

3. The nucleic acid detection cartridge according to claim 1, characterized in that, The nucleic acid testing kit also includes a base (1), which is connected to the lower side of the kit body (2). The card holder body (2) also includes a rotary valve mating part (230), which is made of elastic material and integrally formed on the lower side of the compartment body (200), forming a rotary valve cavity between itself and the base (1). The rotary valve (5) is disposed in the rotary valve cavity and is sealed to the rotary valve mating part (230); and / or The card box body (2) also includes a mounting part (220), which is made of elastic material and integrally formed on the front side wall of the compartment part (200). The mounting part (220) is provided with a through hole for connecting the first flow channel (50) and the amplification cavity (40) of the amplification chip (4). The mounting part (220) is used to seal with the amplification chip (4).

4. The nucleic acid detection cartridge according to claim 3, characterized in that, The valve mating part (230) and / or the mounting part (220) are made of TPE, TPU or liquid silicone material; and / or The compartment body (200) is made of PP, COC, or PC material; and / or The valve mating part (230) and the housing part (200) are injection molded in two colors, and / or the mounting part (220) and the housing part (200) are injection molded in two colors; and / or The surface of the valve mating part (230) forming the valve cavity is fitted with the shape of the valve (5).

5. The nucleic acid detection cartridge according to claim 3, characterized in that, The base (1) includes a side limiting structure (11) located on the front side of the mounting part (220), and a slot (220a) is formed between the side limiting structure (11) and the mounting part (220). The slot (220a) is configured to accommodate a connector (43) located on the rear side of the amplification chip (4), and the mounting part (220) is configured to seal with the connector (43).

6. The nucleic acid detection cartridge according to claim 5, characterized in that, The rear surface of the side limiting structure (11) includes a first inclined surface (111), which forms an acute angle with the vertical direction (A) and gradually slopes backward from top to bottom. The first inclined surface (111) is configured to match the shape of a second inclined surface (431) provided on the front surface of the connector (43).

7. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The card holder body (2) includes multiple drainage holes (2a), which are correspondingly arranged with the multiple receiving cavities and can communicate with the first flow channel (50). The drainage hole (2a) includes a narrowing section whose diameter gradually decreases from top to bottom; and / or At least one of the receiving cavities has a bottom wall comprising an inclined bottom surface that slopes downward from the bottom end of the partition wall (218) surrounding the receiving cavity to the drainage hole (2a).

8. The nucleic acid detection kit according to any one of claims 1-6, characterized in that, The plurality of chambers further includes a first air chamber (262), which is separated from the sample chamber (291) by the partition wall (218); A first air passage (2b) is opened at the top of the partition wall (218) that separates the first air chamber (262) and the sample chamber (291), and the first air passage (2b) connects the sample chamber (291) and the first air chamber (262).

9. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The plurality of accommodating cavities also include a waste liquid cavity (292), which is separated from the extraction cavity (20) by the partition wall (218); A second vent (2c) is provided at the top of the partition wall (218) that separates the waste liquid chamber (292) and the extraction chamber (20), and the second vent (2c) connects the waste liquid chamber (292) and the extraction chamber (20).

10. The nucleic acid detection cartridge according to claim 9, characterized in that, The waste liquid chamber (292) is further provided with a partition wall (219) on its wall, which divides the waste liquid chamber (292) into a waste liquid sub-chamber (2921) and a drying sub-chamber (2922). The drying sub-chamber (2922) is connected to the extraction chamber (20) through the second air outlet (2c). The first flow channel (50) is configured to selectively connect the transfer chamber (201) and the waste liquid sub-chamber (2921). A desiccant is placed in the drying sub-chamber (2922).

11. The nucleic acid detection cartridge according to claim 10, characterized in that, The partition wall (219) is disposed on the partition wall (218) separating the waste liquid chamber (292) and the extraction chamber (20); and / or The top of the partition wall (219) has a third vent hole (2d), which connects the waste liquid chamber (2921) and the drying chamber (2922).

12. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The plurality of chambers further includes a second air chamber (261), which is configured to keep the air pressure value in the amplification chamber (40) within a preset air pressure range; The rotary valve (5) further includes a second flow channel (51), which is configured to connect the amplification chamber (40) and the second gas chamber (261) when the first flow channel (50) connects the intermediate transfer chamber (201) and the amplification chamber (40).

13. The nucleic acid detection cartridge according to claim 12, characterized in that, The chamber (200) further includes a gas flow channel (200b), the amplification chamber (40) is connected to the gas flow channel (200b), and the second flow channel (51) is configured to connect the amplification chamber (40) and the second gas chamber (261) through the gas flow channel (200b) when the first flow channel (50) connects the transfer chamber (201) and the amplification chamber (40).

14. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The card box body (2) includes a first welding part (271), which is provided along the top of the outer wall of the compartment part (200) and / or the top of the partition wall (218); The nucleic acid test kit also includes a cover (3), which is connected above the main body (2) of the kit to close at least a portion of the multiple chambers. The cover (3) includes a second weld (321), which is located at the bottom end of the cover (3) and is configured to be embedded and welded together with the first weld (271).

15. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The nucleic acid testing cartridge also includes a cover (3), which is connected to the upper part of the cartridge body (2) to close at least a portion of the multiple chambers. The cover (3) includes: The lower cover (32) is sealed to the top of the outer wall of the compartment (200) and the top of the plurality of partition walls (218), and includes a plurality of lower cover openings (322) that communicate with a portion of the plurality of chambers, the portion of the chambers including the sample chamber (291) and the transfer chamber (201). A sealing film (3a), connected above the lower cover (32), includes two sealing film openings (30a) respectively opposite to the sample chamber (291) and the transfer chamber (201), for sealing the lower cover openings (322) of the plurality of lower cover openings (322) that correspond to the partial chambers other than the sample chamber (291) and the transfer chamber (201); and The upper cover (31) is movably connected to the lower cover (32) and has a snap-fit ​​state covering the lower cover (32) and the cover sealing film (3a) and an open state exposing the lower cover (32) and the cover sealing film (3a). It includes a sealing part (311) and an upper cover opening (312). In the snap-fit ​​state, the sealing part (311) is opposite to the sample cavity (291) and is configured to block the lower cover opening (322) and the sealing film opening (30a) corresponding to the sample cavity (291). The upper cover opening (312) is opposite to the transfer cavity (201) and the sealing film opening (30a) corresponding to the transfer cavity (201).

16. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The amplification chip (4) further includes a liquid passage (401) for connecting the first flow channel (50) and the amplification cavity (40) and an air passage (42) for keeping the air pressure value in the amplification cavity (40) within a preset air pressure range. The liquid passage (401) includes a large diameter section (4011) away from the amplification cavity (40), a small diameter section (4012) closer to the amplification cavity (40) with a diameter smaller than that of the large diameter section (4011), and a transition section (4013) connecting the large diameter section (4011) and the small diameter section (4012) where the diameter gradually decreases from the large diameter section (4011) to the small diameter section (4012).

17. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The plurality of reagent chambers includes a plurality of cleaning chambers, each pre-stored with a different cleaning solution; and / or The plurality of reagent chambers include the plurality of lyophilized bulb chambers.

18. The nucleic acid detection cartridge according to any one of claims 1-6, characterized in that, The sample cavity (291) is provided with a drainage hole (2a) at the bottom, and the drainage hole (2a) can communicate with the first flow channel (50); The sample chamber (291) is provided with a filter element (2910) extending along the vertical direction (A), and the bottom of the filter element (2910) is arranged opposite to the drainage hole (2a) in the sample chamber (291) along the vertical direction (A).

19. The nucleic acid detection kit according to any one of claims 1-6, characterized in that, The nucleic acid test kit also includes at least one storage container (7), which is movably located in one of the reagent chambers along the vertical direction (A) so as to be in a first position and a second position in the reagent chamber; The card box body (2) includes a spike, which is located at the bottom of the reagent chamber corresponding to the storage container (7). At the first position of the storage container (7), the spike is spaced from the storage container (7) along the vertical direction (A). At the second position of the storage container (7), the spike pierces the storage container (7).

20. The nucleic acid detection cartridge according to claim 19, characterized in that, The storage container (7) includes a sealing portion (73) configured to seal the gap between the storage container (7) and the spike portion.

21. The nucleic acid detection cartridge according to claim 20, characterized in that, The sealing part (73) includes at least one sealing ring disposed at the bottom of the storage container (7) and arranged circumferentially around the spike.

22. The nucleic acid detection cartridge according to claim 19, characterized in that, The nucleic acid test kit also includes a cover (3), which is connected to the upper side of the kit body (2) to close at least a portion of the multiple chambers. The cover (3) includes a first limiting part. The storage container (7) includes a second limiting part. At the first position of the storage container (7), the second limiting part cooperates with the first limiting part to restrict the movement of the storage container (7) in the vertical direction (A).

23. The nucleic acid detection cartridge according to claim 22, characterized in that, The first limiting part includes an elastic claw (34); The second limiting part includes a limiting ring (71), which is disposed on the outer periphery of the storage container (7). At the first position of the storage container (7), the elastic claw (34) is engaged with the bottom of the limiting ring (71). At the second position of the storage container (7), the elastic claw (34) is located above the storage container (7).

24. The nucleic acid detection cartridge according to claim 22, characterized in that, A third limiting part (27a) is provided in the reagent chamber corresponding to the storage container (7). In the second position, the third limiting part (27a) restricts the storage container (7) from moving downward.

25. The nucleic acid detection cartridge according to any one of claims 1-7, characterized in that, Below the extraction cavity (20) is an ultrasonic mating part (250) for cooperating with an ultrasonic device (700), and the bottom surface of the ultrasonic mating part (250) includes a flat surface or a downwardly protruding arcuate surface (20a).

26. A nucleic acid detection system, characterized in that, The nucleic acid detection cartridge includes any one of claims 1-25.

27. A sample processing method based on the nucleic acid detection cartridge according to any one of claims 1-25, characterized in that, include: Step S10: Add the sample to be tested into the sample cavity (291); Step S20: The sample to be tested is lysed into the required lysate within the extraction chamber (20); Step S30: The cleaning solution in the cleaning chamber is transported to the extraction chamber (20) to clean the lysate, and the purification medium in the extraction chamber (20) is used to adsorb nucleic acid to form a lysate suspension and the waste liquid after the nucleic acid adsorption is completed is discharged. Step S40: The eluent in the elution chamber (25) is transported to the extraction chamber (20) to elute the adsorbate adsorbed by the purification medium to form an elution product; Step S50: The elution product is delivered to one of the lyophilized bulb chambers to rehydrate the lyophilized bulb therein, forming the first amplification reaction solution; and Step S60: The first amplification reaction solution is delivered to the amplification chamber (40), and a PCR amplification reaction is performed in the amplification chamber (40) to form the first amplification product.

28. The sample processing method according to claim 27, characterized in that, The plurality of reagent chambers includes two lyophilized bulb chambers, wherein, after step S60, the sample processing method further includes: Step S70: The first amplification product formed in step S60 is delivered to the elution chamber (25), and the first amplification product is diluted with the elution buffer in the elution chamber (25); Step S80: The diluted first amplification product is delivered to the other of the two lyophilized bulb chambers to re-dissolve the lyophilized bulb therein to form a second amplification reaction solution; Step S90: The second amplification reaction solution is delivered to the amplification chamber (40), and a PCR amplification reaction is performed in the amplification chamber (40) to form the second amplification product.

29. The sample processing method according to claim 27 or 28, characterized in that, Step S10 includes: adding the sample or a sample mixture of the sample and the preservative into the sample chamber (291) as the sample to be tested; Step S20 includes: transporting the sample to be tested in the sample chamber (291) and the lysis buffer in the lysis chamber (21) to the extraction chamber (20) to lyse the sample to be tested, using the purification medium in the extraction chamber (20) to adsorb nucleic acid and discharging the waste liquid after nucleic acid adsorption.

30. The sample processing method according to claim 27 or 28, characterized in that, Step S10 includes: processing the sample with a lysis buffer other than the nucleic acid detection cartridge to form the sample to be tested, and adding the sample to be tested into the sample chamber (291); Step S20 includes: transferring the sample to be tested from the sample chamber (291) to the extraction chamber (20) in multiple transfers, performing nucleic acid adsorption after each transfer and discharging the waste liquid after nucleic acid adsorption is completed, until all samples to be tested are processed.

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