Nucleic acid extraction device

Abstract

The utility model relates to the technical field of nucleic acid extraction, and in particular relates to a nucleic acid extraction device which comprises a sample adding part, a first extraction part and a second extraction part, the connecting part is connected with the sample adding part, a puncture part is formed at the first end of the connecting part, and a first connecting cavity is formed below the puncture part; when the sample adding part and the connecting part are in a first connecting state, a distance exists between the puncture end and the first containing cavity, the sample adding part and the connecting part are converted into a second connecting state from the first connecting state, and the puncture end punctures the wall of the first containing cavity so as to guide liquid to enter the first connecting cavity from the side face of the puncture end; the reaction part is connected with the connecting part, and a second containing cavity is formed in the reaction part. The utility model provides an integrated nucleic acid extraction device which has the characteristic of simplicity and convenience in operation.

Description

Technical Field

[0001] This utility model relates to the field of nucleic acid extraction for molecular biological detection, and specifically to a nucleic acid extraction device. Background Technology

[0002] Existing nucleic acid extraction methods commonly include magnetic bead extraction, centrifugal column extraction, and ultrasonic extraction. However, these methods require large-scale equipment (such as centrifuges, magnetically driven equipment, etc.) and have long extraction times. Manual extraction methods include phenol extraction, alkaline lysis, CTAB extraction, and boiling.

[0003] For example, patent application CN202311677304.7 discloses a sample processing method, which involves mixing the sample to be tested with a direct amplification sample processing solution and then sonicating it to obtain a sample nucleic acid solution.

[0004] For example, patent application CN118234843A describes an automated nucleic acid extraction kit and an automated nucleic acid extraction system including the kit. The kit has a housing comprising a sample port, a cell processing chamber, a washing chamber, a filter assembly including a filter element, and a diverter valve having a first reversible sealed output and a second reversible sealed output. The sample port and cell processing chamber, the cell processing chamber and filter assembly, and the washing chamber and filter assembly communicate via unidirectional fluid channels. The filter assembly communicates fluidly with the diverter valve and (i) communicates with a waste liquid pipeline when the diverter valve is biased to the first reversible sealed output, and (ii) communicates with a pathogen nucleic acid pipeline when the diverter valve is biased to the second reversible sealed output. However, this automated nucleic acid extraction system has a very complex structure, and its processing difficulty and application cost are very high, making it unsuitable for applications such as large-scale nucleic acid extraction.

[0005] For example, patent application CN200980133630.3 discloses a disposable device for automated biological sample preparation. This disposable device has a connection mechanism for connecting to a sealed sonication tube, and a filter membrane capture unit for capturing and processing the sample without requiring other tubes. The operation of this device is also very cumbersome, requiring the sonication tube to be placed in an ultrasonic disruptor and the stopper spacer removed during automated operation. Furthermore, the disposable device requires external equipment such as buffer bottles, waste bottles, washing bottles, elution bottles, and PCR reagent bottles to complete nucleic acid extraction. Depending on the target substance to be separated, other types of buffers and reagents can be used. The tube is pre-loaded with measured sample preparation reagents for a single sample preparation operation. The bottle can be sealed with a membrane at the top to prevent liquid evaporation during storage.

[0006] Therefore, both the automated and manual extraction equipment mentioned above suffer from cumbersome operation. With the development of clinical molecular diagnostics, pathogen nucleic acid extraction followed by PCR amplification and identification can serve as a clinical reference, and molecular biology techniques play a crucial role in disease diagnosis. The quality of the nucleic acid sample is critical to subsequent results; when the quality of the nucleic acid fails to meet requirements, it will affect the accuracy and detection limit of subsequent test results. Furthermore, the sample collection and transportation process, extraction methods, and efficiency all influence nucleic acid quality. In addition, general nucleic acid extraction requires highly skilled instruments and personnel, and the entire extraction process is relatively lengthy.

[0007] Therefore, there is an urgent need for a device and method that can rapidly extract nucleic acids. Summary of the Invention

[0008] The purpose of this invention is to provide a nucleic acid extraction device and method that partially solves or alleviates the above-mentioned deficiencies in the prior art and simplifies the nucleic acid extraction operation.

[0009] To solve the aforementioned technical problems, the present invention specifically adopts the following technical solution:

[0010] A nucleic acid extraction device, comprising:

[0011] The sample dispensing section is provided with a first receiving cavity, which is used to contain a first sample liquid.

[0012] A connecting portion is connected to the sample application portion. A puncture portion is formed at the first end of the connecting portion. The puncture portion has a puncture end capable of piercing the wall of the first receiving cavity. A first connecting cavity is formed below the puncture portion. The sample application portion and the connecting portion are connected in a rotatable manner. When the sample application portion and the connecting portion are in a first connected state, there is a gap between the puncture end and the first receiving cavity. When the sample application portion rotates along a preset first direction, the sample application portion and the connecting portion change from the first connected state to a second connected state. The puncture end pierces the wall of the first receiving cavity to guide the first sample liquid into the first connecting cavity from the side of the puncture end.

[0013] The reaction section is connected to the connecting section, and the reaction section has a second receiving cavity for containing a second sample liquid.

[0014] In some embodiments, the connecting portion has a first surface with a limiting protrusion, and the sample feeding portion has a second surface with a limiting path, the limiting path comprising: a first limiting segment and a second limiting segment that intersect; wherein...

[0015] When the sample feeding part and the connecting part are in the first connection state, the limiting protrusion is engaged in the first limiting segment; when the limiting protrusion moves from the first limiting segment into the second limiting segment, the sample feeding part and the connecting part change from the first connection state to the second connection state.

[0016] In some embodiments, a third limiting segment connected to the second limiting segment is further provided along the direction from the second end to the first end of the connecting portion, and the third limiting segment is intersecting with the second limiting segment; correspondingly, when the sample application portion and the connecting portion are in the second connecting state from the first connecting state, a gap is formed between the puncture end and the opening of the sample application portion;

[0017] When the limiting protrusion moves from the second limiting segment to the third limiting segment, the sample application part and the connecting part change from the second connection state to the third connection state, and the distance between the puncture end and the opening decreases so that the puncture end is adjacent to the opening.

[0018] In some embodiments, the diameter of the puncture portion gradually increases in the direction gradually away from the puncture end, and a plurality of drainage channels are provided at intervals on the side of the puncture portion to guide the first sample liquid from the puncture end into the first connecting cavity.

[0019] In some embodiments, the puncture portion and the first wall of the connecting portion are connected by a curved second wall, the second wall being recessed toward the side away from the puncture end; wherein the first wall, the second wall and the wall of the puncture portion cooperate to form a curved drainage surface, the drainage surface being able to restrict the flow of liquid to the connection between the sample application portion and the connecting portion.

[0020] In some embodiments, the first sample liquid is an acidic liquid.

[0021] In some embodiments, the second sample liquid is a lysis buffer.

[0022] In some embodiments, the second end of the reaction section is recessed inward to form a recessed region.

[0023] In some embodiments, the recessed area is coated with a solid substance for lubrication.

[0024] In some embodiments, the reaction part and the connecting part are connected by a thread.

[0025] Beneficial technical effects:

[0026] This invention provides an integrated nucleic acid extraction device, enabling testing personnel to extract nucleic acids from samples such as whole blood. Furthermore, the extracted nucleic acids can be stably preserved and transported directly within the device, thereby significantly reducing the labor intensity during batch nucleic acid extraction and substantially decreasing the workload of testing personnel.

[0027] Specifically, the cross-design of the multi-stage limiting sections assists users in quickly puncturing the first receiving cavity to release the first sample liquid by rotating the connecting part or the sample dispensing part. Simultaneously, this multi-stage limiting design maintains the stability of the nucleic acid extraction device during transportation. For example, when the nucleic acid extraction device is subjected to bumps or impacts during transportation, the limiting function of the limiting sections prevents relative rotation between the connecting part and the sample dispensing part caused by the bumps, thus avoiding premature release of the first sample liquid.

[0028] This novel nucleic acid extraction device only requires the use of ultrasonic equipment to complete on-site nucleic acid release, facilitating rapid on-site operation by testing personnel and minimizing limitations on equipment and location. This extraction device can be widely applied in large-scale testing scenarios such as on-site sampling and remote testing. It is worth noting that in these large-scale testing scenarios, the number of subjects is numerous, making sampling and testing extremely demanding and requiring very high efficiency. Furthermore, such large-scale testing activities often involve lengthy on-site sampling and remote testing. Attached Figure Description

[0029] To more clearly illustrate the technical solutions in the embodiments of this utility model or the prior art, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. The elements or parts in the drawings are not necessarily drawn to scale. Obviously, the drawings described below are some embodiments of this utility model. For those skilled in the art, other drawings can be obtained based on these drawings without any creative effort.

[0030] Figure 1 This is a cross-sectional structural diagram of the device in an exemplary embodiment of the present invention;

[0031] Figure 2 This is a schematic diagram of the device in an exemplary embodiment of the present invention;

[0032] Figure 3 This is an exploded view of the components of the device in an exemplary embodiment of the present invention;

[0033] Figure 4This is a schematic diagram of the connection portion of the device in an exemplary embodiment of the present invention;

[0034] Figure 5 This is a schematic diagram of the sample feeding section of the device in an exemplary embodiment of the present invention;

[0035] Figure 6 This is a schematic diagram of the limiting path of the device in an exemplary embodiment of the present invention;

[0036] Figure 7 This is a schematic diagram of the device in another exemplary embodiment of the present invention.

[0037] Summary of reference numerals in the attached drawings: Sample feeding section 10, first receiving cavity 11, cover 12, second surface 13, limiting path 14, first limiting segment 141, second limiting segment 142, third limiting segment 143, fourth limiting segment 144, first space 15, second space 16; connecting part 20, puncture part 21, drainage channel 211, first connecting cavity 22, first surface 23, limiting protrusion 24, second connecting cavity 25; reaction part 30, second receiving cavity 31, recessed area 32, spring 40. Detailed Implementation

[0038] To make the objectives, technical solutions, and advantages of the embodiments of this utility model clearer, the technical solutions of the embodiments of this utility model will be clearly and completely described below with reference to the accompanying drawings. Obviously, the described embodiments are only some, not all, of the embodiments of this utility model. All other embodiments obtained by those skilled in the art based on the embodiments of this utility model without creative effort are within the scope of protection of this utility model.

[0039] In this document, suffixes such as "module," "part," or "unit" used to denote elements are used only for the purpose of illustrative purposes and have no specific meaning in themselves. Therefore, "module," "part," or "unit" can be used interchangeably.

[0040] In this document, the terms "upper," "lower," "inner," "outer," "front," "rear," "one end," and "the other end," etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are used only for the convenience of describing this utility model and simplifying the description, and 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. Therefore, they should not be construed as limitations on this utility model. Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance.

[0041] In this document, unless otherwise explicitly specified and limited, the terms "installed," "equipped with," and "connected," etc., should be interpreted broadly. For example, "connected" can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection, a direct connection, or an indirect connection through an intermediate medium; it can be a connection within two components. Those skilled in the art can understand the specific meaning of the above terms in this utility model based on the specific circumstances.

[0042] In this document, "and / or" includes any and all combinations of one or more of the listed related items.

[0043] In this article, "multiple" means two or more, that is, it includes two, three, four, five, etc.

[0044] As used in this specification, the term "about" typically means + / -5% of the value, more typically + / -4% of the value, more typically + / -3% of the value, more typically + / -2% of the value, even more typically + / -1% of the value, and even more typically + / -0.5% of the value.

[0045] In this specification, certain embodiments may be disclosed in a range-bound format. It should be understood that this "range-bound" description is merely for convenience and brevity and should not be construed as a rigid limitation on the disclosed range. Therefore, the description of a range should be considered as having specifically disclosed all possible subranges and the individual numerical values ​​within those ranges. For example, a description of the range 1-6 should be considered as having specifically disclosed subranges such as from 1 to 3, from 1 to 4, from 1 to 5, from 2 to 4, from 2 to 6, from 3 to 6, etc., and the individual numbers within those ranges, such as 1, 2, 3, 4, 5, and 6. This rule applies regardless of the breadth of the range.

[0046] This specification uses the terms "patient" or "object" to describe animals, such as mammals, preferably humans or domesticated animals. In any aspect or specific embodiment described in this disclosure, the object or patient may also be a primate. In any aspect or specific embodiment described in this disclosure, the patient or object is a human. In any aspect or specific embodiment described in this disclosure, the patient or object is livestock, such as cattle, sheep, goats, dairy cows, pigs, etc.; or domesticated animals, such as dogs, fish, guinea pigs, and cats. In any aspect or specific embodiment described in this disclosure, the object is a rodent (e.g., mouse, rat, hamster), rabbit, primate, non-human primate, or pig, such as a purebred pig.

[0047] In this article, radial refers to the direction passing through the axis (or axial line) within the radial plane. The radial plane is a plane that is perpendicular (or approximately perpendicular) to the axis.

[0048] In this article, see Figure 6 As shown, when the connecting part rotates continuously in the first direction, the limiting protrusion moves continuously in the limiting path. The two ends of the limiting segment can be referred to as the first end and the second end of the limiting segment along the direction of movement. The first direction can also be called the unlocking direction. When the connecting part moves in the first direction, the distance between the puncture end of the connecting part and the opening of the sample application part gradually decreases. Correspondingly, when the connecting part rotates continuously in the second direction (opposite to the first direction), the distance between the puncture end of the connecting part and the opening of the sample application part will gradually increase.

[0049] See Figures 1-7 As shown, this utility model provides a nucleic acid extraction device.

[0050] Preferably, the nucleic acid extraction device of this invention includes:

[0051] The sample dispensing section 10 is provided with a first receiving cavity 11, which is used to contain a first sample liquid (e.g., HCl).

[0052] A connecting part 20 is connected to the sample application part 10. A puncture part 21 is formed at the first end of the connecting part. The puncture part has a puncture end that can puncture the wall of the first receiving cavity. A first connecting cavity 22 is formed below the puncture part (i.e. on the side away from the puncture end).

[0053] The connecting part 20 has a first surface 23, on which a limiting protrusion 24 is provided; the sample feeding part 10 has a second surface 13, on which a limiting path 14 is provided; the limiting path 14 includes a first limiting segment 141 of a certain length and a second limiting segment 142 of a certain length arranged sequentially along the direction from the second end to the first end of the sample feeding part, and the first limiting segment and the second limiting segment 142 are intersecting.

[0054] When the sample application part and the connecting part are in the first connection state, the limiting protrusion is engaged in the first limiting segment. At this time, there is a certain gap between the puncture end and the first receiving cavity (specifically, the lower surface of the first receiving cavity). When the limiting protrusion moves from the first limiting segment into the second limiting segment, the sample application part and the connecting part change from the first connection state to the second connection state. At this time, the gap between the puncture end and the first receiving cavity gradually decreases, and the puncture end punctures the wall of the first receiving cavity to guide the first sample liquid from the side of the puncture end into the first connecting cavity 22.

[0055] The reaction section 30 is connected to the connecting section, and the reaction section has a second receiving cavity 31 for containing a second sample liquid (e.g., NaOH). Specifically, the first connecting cavity 22 is connected to the second receiving cavity 31.

[0056] In some embodiments, when it is necessary to store the released nucleic acid for a long time, the first end of the sample dispensing part has an openable cover 12, which can provide a sealing and protection function for the nucleic acid when the cover is closed.

[0057] Of course, in some scenarios where on-site sampling and testing are conducted, the sample dispensing section only needs to have an opening that allows liquid to flow out.

[0058] Preferably, see Figure 6 As shown, the first limiting segment 141 and the second limiting segment 142 are intersecting, so that when the limiting protrusion 24 is engaged in the first limiting segment 141, the wall of the first limiting segment 141 (which has a certain length) can limit the limiting protrusion 24, so as to prevent the limiting protrusion 24 from making improper displacement and affecting the stability of the internal liquid.

[0059] For example, the overlapping arrangement of the limiting sections helps users quickly puncture the first receiving cavity to release the first sample liquid by rotating the connecting part or the sample dispensing part. Simultaneously, this multi-segment limiting design also maintains the stability of the nucleic acid extraction device during transportation. For instance, when the nucleic acid extraction device is subjected to bumps or impacts during transportation, the limiting effect of the limiting sections can prevent relative rotation between the connecting part and the sample dispensing part caused by the bumps, thus avoiding premature release of the first sample liquid.

[0060] Furthermore, the second limiting segment can be arranged in a direction parallel or approximately parallel to the radial plane. Thus, when the limiting protrusion is stuck in the middle position of the second limiting segment with a certain length, the second limiting segment can keep the puncture segment and the first receiving cavity in a specific connection state, so as to facilitate the liquid to smoothly enter the first connecting cavity under the guidance of the puncture end, and then enter the reaction section.

[0061] For example, in some embodiments, the limiting path can be a slit that runs through the wall of the sample feeding section to limit the limiting protrusion.

[0062] In some embodiments, the spring 40 is further included, wherein when the sample feeding part is connected to the connecting part, and when the limiting protrusion is engaged with the first limiting segment, the two ends of the spring abut against the end faces of the connecting part and the sample feeding part respectively.

[0063] This invention provides a nucleic acid extraction device that integrates multiple functions such as sample storage and processing. The sample dispensing part and the connecting part of the extraction device cooperate with the spring through a multi-level limiting and locking function to provide multiple stable connection states (such as the first connection state and the second connection state) to adapt to the operational needs at different stages.

[0064] When the user rotates the connecting part (or the sample feeding part), the spring, which cooperates with the multi-segment limiting section, provides a certain degree of feedback force to the user. Specifically, when the connecting part rotates relative to the sample feeding part in the first direction (i.e., the unlocking direction), the distance between the two end faces gradually decreases. At this time, the spring contracts, and the force exerted by the spring on the end faces (equivalent to the feedback force) also gradually increases. Thus, the user can comprehensively judge the rotation operation status at this time by the magnitude of the force perceived by their hand.

[0065] At the same time, the spring also serves as a reset function. Specifically, when the user applies an external force to the connecting part to rotate it, the piercing end punctures the first receiving cavity (at which point the spring is compressed). When the user removes the applied external force, the spring automatically extends under the reset function, thereby causing the limiting protrusion to return to its initial position under the reset function of the spring.

[0066] Preferably, see Figure 1 As shown, the wall of the sample feeding part 10 has a protruding end face along its radial direction (or approximately radial direction), and the wall of the connecting part 20 also has a protruding end face along its radial direction (or approximately radial direction). The two end faces are arranged opposite to each other, and a spring 40 is arranged between the two end faces. The spring is sleeved on the second surface of the sample feeding part and the first surface of the connecting part.

[0067] In this embodiment, when the user rotates the connecting part (or the sample feeding part), the spring can provide a certain degree of feedback force to the user. Specifically, when the connecting part rotates relative to the sample feeding part along the first direction (i.e., the direction used to pierce the first receiving cavity, also known as the unlocking direction), the distance between the two end faces gradually decreases. At this time, the spring contracts, and the force exerted by the spring on the end faces (equivalent to the feedback force) also gradually increases. Thus, the user can comprehensively judge the rotation operation state at this time by the magnitude of the force perceived by their hand.

[0068] Furthermore, the spring also serves a reset function. Specifically, when the user applies an external force to the connecting part to rotate it, causing the limiting protrusion 24 to move from the first end of the first limiting segment 141 to the second end, the piercing end also pierces the first receiving cavity (at which point the spring is compressed); correspondingly, when the user cancels the applied external force, the spring can automatically extend under the reset function, thereby causing the limiting protrusion 24 to return to the second end of the first limiting segment 141 under the reset function of the spring.

[0069] In some embodiments, see Figure 6 As shown, a third limiting segment 143 connected to the second limiting segment is also provided along the direction from the second end to the first end of the connecting portion, and the third limiting segment is intersecting with the second limiting segment.

[0070] Correspondingly, when the sample application part and the connecting part are in the second connection state, a gap is formed between the puncture end and the opening of the sample application part (this can also prevent excessive liquid from accumulating in the opening area to a certain extent and prevent liquid leakage at the opening); when the limiting protrusion moves from the second limiting segment to the third limiting segment, the sample application part and the connecting part change from the second connection state to the third connection state, and the gap between the puncture end and the opening decreases so that the puncture end is adjacent to the opening.

[0071] In some embodiments, the diameter of the puncture portion gradually increases in the direction gradually away from the puncture end (in other words, the diameter of the puncture portion gradually increases in the direction from its first end to its second end), and a plurality of drainage channels 211 are provided at intervals on the side of the puncture portion to guide the first sample liquid from the puncture end into the first connecting cavity.

[0072] In some embodiments, see Figure 7 As shown, the puncture portion 21 is connected to the first wall 25a (i.e., the wall of the second connecting cavity 25) of the connecting portion 20 by a curved second wall 21a, the second wall 21a being recessed toward the side away from the puncture end; the first wall is in close contact with the inner surface of the sample application portion;

[0073] For example, see Figure 1 As shown, the cavity within the connecting portion 20 is divided into a first connecting cavity 22 and a second connecting cavity 25 by the puncture portion 21, and both are used to connect to the reaction portion 30 and the sample application portion 10, respectively. Specifically, the first wall of the second connecting cavity 25 is formed by extending from the second end of the puncture portion 21 away from the first connecting cavity 22; thus, the first wall, the second wall, and the outer wall surface of the puncture portion 21 cooperate to form a curved drainage surface. The drainage surface can, on the one hand, guide the liquid directly through the drainage surface into the lower first connecting cavity 22 during the puncture process (avoiding excessive liquid entering the junction between the second space 16 and the connecting portion to prevent leakage); on the other hand, when the device is inverted for sample inversion, the drainage surface can still play a similar leak-proof role.

[0074] Furthermore, in some embodiments, see Figure 6 As shown, along the direction from the second end to the first end of the connecting portion (e.g.) Figure 6 A fourth limiting segment 144, having a certain length and connected to the third limiting segment 143, is also provided on the central section (as shown in F). The fourth limiting segments 144 connected to the third limiting segment 143 are also arranged intersectingly. Preferably, the fourth limiting segment 144 can also be arranged in a direction parallel or approximately parallel to the radial plane, thereby ensuring that when the limiting protrusion 24 is engaged with the fourth limiting segment, the device remains in a state adjacent to the opening at the puncture end.

[0075] Furthermore, after the nucleic acid extraction device has completed the extraction of nucleic acid, the device can be inverted so that the liquid inside the device can be released through the opening of the sample dispensing section (i.e., inverting the sample).

[0076] In this embodiment, when the nucleic acid extraction device is inverted, the multi-stage limiting segment design assists the user in ensuring that the puncture tip is adjacent to the opening of the sample application section (e.g., the puncture tip can even be flush with or protrude from the opening). Thus, under the drainage effect of the puncture tip, the liquid can flow directly outwards without excessively flowing to the inner surface of the opening (thereby, to some extent, preventing leakage between the connection and the sample application section). Therefore, when the collected nucleic acid involves different testing items and requires multiple inversions, the multi-stage locking scheme combined with the inverted puncture tip can reduce potential contamination during the inversion process.

[0077] In this embodiment, a multi-level locking scheme is formed by the relative arrangement of multiple limiting segments in the direction. This multi-level locking scheme provides multiple connection states for the nucleic acid extraction device. These multiple connection states can meet the needs of users in different stages such as puncture and sample pouring. Moreover, the operation is simple and can be achieved by simply rotating the connection part continuously.

[0078] In other words, this utility model achieves multi-functional integration (i.e., provides an integrated device) through a multi-level locking scheme between the connecting part and the sample dispensing part. Multiple functions such as sample liquid prediction, nucleic acid release, lysis buffer synthesis, and nucleic acid storage can all be realized through this nucleic acid extraction device.

[0079] Of course, in other embodiments, the limiting path can also be set as a straight path, in which case the connection state between the connecting part and the opening part can be manually controlled by the user.

[0080] Furthermore, in some embodiments, see Figure 4 As shown, the sample feeding section 10 has a first space 15 and a second space 16 formed at both ends of the first receiving cavity 11. The first space has an opening at the end away from the first receiving cavity, and the second space 16 is used to accommodate the puncture section.

[0081] Specifically, see Figure 1 As shown, when the sample feeding part and the connecting part are in the connected state, the inner surface (i.e., the third surface) of the sample feeding part and the outer surface (i.e., the second surface) of the connecting part are closely fitted together.

[0082] In some embodiments, a sealing layer (also referred to as an anti-slip layer) may be provided on the inner surface of the sample feeding part and / or the outer surface of the connecting part. For example, in some embodiments, the sealing layer may be a threaded structure, or the sealing layer may be an elastic material such as rubber, to further improve the sealing performance between the sample feeding part and the connecting part.

[0083] In some embodiments, the first sample liquid is an acidic liquid.

[0084] In some embodiments, the second sample liquid is a lysis buffer. Specifically, the lysis buffer may be an alkaline liquid.

[0085] In some embodiments, the device further includes an ultrasonic device having a mounting position for accommodating the reaction section. When the extraction device is disposed in the ultrasonic device, its spring prevents the ultrasonic action from interfering with the connection between the connecting section and the sample dispensing section, thus preventing premature release of the first sample liquid.

[0086] In some embodiments, the second end of the reaction section 30 is recessed inward to form a recessed region 32, and a solid substance for lubrication is coated on the recessed region 32.

[0087] Correspondingly, the use of solid materials can reduce the loss of ultrasound waves during propagation to some extent.

[0088] Furthermore, in some embodiments, a raised area corresponding to the recessed area is formed on the mounting position of the ultrasonic device, so as to achieve efficient transmission of ultrasonic waves through the cooperation of the recessed area and the raised area, and at the same time, it can also fix the reaction part.

[0089] In some embodiments, the first end of the reaction part 30 and the second end of the connecting part 20 are rotatably connected by a thread.

[0090] In some embodiments, the sample feeding part is connected to the cover body by threads.

[0091] Alternatively, in some embodiments, the cover may be a rubber cover with elasticity, which can snap into the opening above the sample dispensing part to seal the opening.

[0092] In some embodiments, the upper and lower surfaces of the first receiving cavity 11 are formed of a sealing film (such as a thin film, aluminum foil film, etc.) so that the puncture end can puncture.

[0093] In some embodiments, the connecting part can be directly removed to pour out the liquid in the reaction part, so the sample feeding part does not need to be provided with an opening for pouring out the sample.

[0094] To more clearly illustrate the technical solution and beneficial effects of the nucleic acid extraction device of this utility model, the extraction methods will be explained below using whole blood (equivalent to the third sample), HCl (equivalent to the first sample liquid), and NaOH (equivalent to the second sample liquid) as examples:

[0095] This utility model also provides a nucleic acid extraction method, including the following steps:

[0096] S101 uses the nucleic acid extraction device described in any of the embodiments herein;

[0097] S102 Add a third sample (such as whole blood) to be tested into the reaction section, and the reaction section is also provided with a second sample liquid;

[0098] For example, during the transportation of a nucleic acid extraction device, the sample loading section, the connecting section, and the reaction section can be connected sequentially. During the nucleic acid extraction operation, the connecting section 20 is first removed so that the sample to be tested (equivalent to a third sample, such as whole blood, serum, swabs, feces, urine, sputum, or other different types of samples) can be added through the opening at the first end of the reaction section 30.

[0099] S103 uses an ultrasonic device to ultrasonically treat the reaction section, and the second sample liquid (such as NaOH) under the action of ultrasound causes the third sample to release nucleic acid;

[0100] In this embodiment, low-frequency ultrasound is used to induce vibrations and release nucleic acids while maintaining their integrity. Under ultrasonic treatment, NaOH promotes the rapid release of nucleic acids from whole blood.

[0101] Further, the first unlocking operation (or puncture operation) is performed, for example, by performing step S104, which causes the sample application part and the connecting part to change from a first connection state to a second connection state, so that the first sample liquid (such as HCl) in the connecting part enters the first connecting cavity from the side of the puncture end, and the first sample liquid and the second liquid sample undergo a neutralization reaction.

[0102] Furthermore, in some embodiments, the method further includes the step of performing a second unlocking operation to complete the sample reversal; specifically, it includes the following steps:

[0103] S105 causes the sample application part and the connecting part to change from a second connection state to a third connection state, so that the puncture end is close to the opening of the sample application part;

[0104] S106 The nucleic acid extraction device is inverted, and the liquid in the reaction section flows out through the opening under the guidance of the puncture end.

[0105] In some embodiments, different ultrasonic conditions may be used for different types of third samples, as illustrated below:

[0106] (1) Oral / nasopharyngeal swabs, vaginal swabs or other swabs, serum samples: ultrasound frequency: 60-100kHz, time: 3-30 seconds;

[0107] (2) Sputum: Ultrasound frequency: 100kHz~300kHz, time: 5-60 seconds;

[0108] The extraction method of this invention combines ultrasound and strong alkali lysis, allowing the direct release of nucleic acids from sputum or whole blood using this device. Furthermore, this integrated structural design allows for direct neutralization of the lysis buffer via a rotating puncture operation, followed by neutralization with HCl after lysis, thus avoiding the influence of NaOH on the PCR reaction (polymerase chain reaction). Therefore, by continuously performing ultrasonic strong alkali lysis and puncture operations to neutralize the lysis buffer on the nucleic acid extraction device, nucleic acid extraction from samples such as whole blood can be completed using this integrated device. The extracted nucleic acids can be stably stored and transported directly within the device, significantly reducing the labor intensity and workload of testing personnel during batch nucleic acid extraction.

[0109] Furthermore, the nucleic acid extraction device of this invention only needs to be used with ultrasonic equipment to complete the on-site nucleic acid release, which is conducive to rapid on-site operation by testing personnel, and the restrictions on the equipment and the site are relatively small.

[0110] For example, the extraction device of this invention can be widely used in large-scale testing scenarios such as on-site sampling and remote testing. It is worth noting that in these large-scale testing scenarios, the number of objects is numerous, making sampling and testing extremely labor-intensive and requiring very high efficiency. Furthermore, such large-scale testing activities often involve long periods of on-site sampling and remote testing.

[0111] For example, it may be necessary to conduct long-term on-site sampling of subjects (such as residents of a community) in areas far from hospitals or testing laboratories, and then transport the samples to the hospital or testing laboratory. Another example is livestock raised in farms in remote areas.

[0112] Transporting samples from the testing site to the laboratory takes time and can encounter various problems, such as sample damage due to improper storage, sample loss leading to reduced quantity, and liquid contamination during transportation. Furthermore, in large-scale testing scenarios, the sampling workload for testing personnel is extremely heavy, resulting in low efficiency in on-site nucleic acid sampling and extraction.

[0113] In response, this invention proposes an integrated nucleic acid extraction device that can quickly perform operations such as ultrasound, strong alkali lysis, and neutralization of the lysis buffer. This nucleic acid extraction device only requires one ultrasound device to complete the extraction of nucleic acid, thus effectively reducing the detection cost in terms of equipment configuration, and also reducing the amount of manual operation required by the testing personnel.

[0114] Furthermore, the nucleic acid extraction device, through its multi-level locking design, can stably store the predicted sample liquid within the device. At the same time, the multi-level locking design allows testing personnel to manually complete operations such as neutralizing the lysis buffer and pouring out the nucleic acid simply by rotating the connecting part, greatly reducing the workload of testing personnel in on-site nucleic acid extraction.

[0115] Furthermore, the multi-level locking design also helps maintain the relative stability between the internal structures of the extraction device during long-distance transportation, ensuring that it can stably store nucleic acids.

[0116] The continuous ultrasound and strong alkali lysis operation method implemented by this nucleic acid extraction device has strong versatility and can be applied to different types of samples such as whole blood, serum, swabs, feces, urine, and sputum.

[0117] It should be noted that, in this document, the terms "comprising," "including," or any other variations thereof are intended to cover non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements includes not only those elements but also other elements not expressly listed, or elements inherent to such a process, method, article, or apparatus. Unless otherwise specified, an element defined by the phrase "comprising one..." does not exclude the presence of other identical elements in the process, method, article, or apparatus that includes that element.

[0118] The embodiments of the present invention have been described above with reference to the accompanying drawings. However, the present invention is not limited to the specific embodiments described above. The specific embodiments described above are merely illustrative and not restrictive. Those skilled in the art can make many other forms under the guidance of the present invention without departing from the spirit and scope of the claims. All of these forms are within the protection scope of the present invention.

Claims

1. A nucleic acid extraction device, characterized in that, include: The sample feeding section (10) is provided with a first receiving cavity (11) for containing a first sample liquid; A connecting part (20) is connected to the sample application part (10). A puncture part (21) is formed at the first end of the connecting part. The puncture part has a puncture end capable of piercing the wall of the first receiving cavity. A first connecting cavity (22) is formed below the puncture part. The sample application part and the connecting part are connected in a rotatable manner. When the sample application part and the connecting part are in a first connection state, there is a gap between the puncture end and the first receiving cavity. When the sample application part rotates along a preset first direction, the sample application part and the connecting part change from the first connection state to a second connection state. The puncture end pierces the wall of the first receiving cavity to guide the first sample liquid into the first connecting cavity from the side of the puncture end. The reaction section (30) is connected to the connecting section and has a second receiving cavity (31) for receiving a second sample liquid.

2. The nucleic acid extraction device according to claim 1, characterized in that, The connecting part (20) has a first surface (23) on which a limiting protrusion (24) is provided; the sample feeding part (10) has a second surface (13) on which a limiting path (14) is provided; the limiting path includes: a first limiting segment and a second limiting segment that are intersected; wherein... When the sample feeding part and the connecting part are in the first connection state, the limiting protrusion is engaged in the first limiting segment; when the limiting protrusion moves from the first limiting segment into the second limiting segment, the sample feeding part and the connecting part change from the first connection state to the second connection state.

3. The nucleic acid extraction device according to claim 2, characterized in that, A third limiting segment (143) connected to the second limiting segment is also provided along the direction from the second end to the first end of the connecting part, and the third limiting segment is intersecting with the second limiting segment; correspondingly, when the sample application part and the connecting part change from the first connection state to the second connection state, a gap is formed between the puncture end and the opening of the sample application part; When the limiting protrusion moves from the second limiting segment to the third limiting segment, the sample application part and the connecting part change from the second connection state to the third connection state, and the distance between the puncture end and the opening decreases so that the puncture end is adjacent to the opening.

4. The nucleic acid extraction device according to claim 1, characterized in that, The diameter of the puncture portion gradually increases in the direction away from the puncture end, and multiple drainage channels are provided at intervals on the side of the puncture portion to guide the first sample liquid from the puncture end into the first connecting cavity.

5. A nucleic acid extraction device according to any one of claims 1-4, characterized in that, The puncture portion and the first wall of the connecting portion are connected by a curved second wall, which is recessed toward the side away from the puncture end; wherein the first wall, the second wall and the wall of the puncture portion (21) cooperate to form a curved drainage surface, which can restrict the flow of liquid to the connection between the sample application portion and the connecting portion.

6. The nucleic acid extraction device according to claim 1, characterized in that, The first sample liquid is an acidic liquid.

7. The nucleic acid extraction device according to claim 1, characterized in that, The second sample liquid is a lysis buffer.

8. The nucleic acid extraction device according to claim 1, characterized in that, The second end of the reaction section (30) is recessed inward to form a recessed area.

9. A nucleic acid extraction device according to claim 8, characterized in that, The recessed area is coated with a solid substance for lubrication.

10. A nucleic acid extraction device according to claim 1, characterized in that, The reaction part (30) and the connecting part are connected by a thread.

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