Nucleic acid extraction device and method

Abstract

The invention relates to the technical field of nucleic acid extraction, in particular to a nucleic acid extraction device and method.The nucleic acid extraction device 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 invention provides an integrated nucleic acid extraction device which has the characteristic of simplicity and convenience in operation.

Description

Technical Field

[0001] The present invention relates to the field of nucleic acid extraction for molecular biology detection, and in particular to a nucleic acid extraction device and method. Background Art

[0002] The existing nucleic acid extraction methods commonly used include magnetic bead extraction, centrifugal column extraction, ultrasonic extraction, etc., but these methods require large-scale equipment (such as centrifugal equipment, magnetic drive equipment, etc.), and the nucleic acid extraction time is relatively long. Manual extraction methods include phenol extraction, alkaline lysis, CTAB extraction, boiling, etc.

[0003] For example, patent application CN202311677304.7 discloses a sample processing method, which mixes the sample to be tested with a direct diffusion sample processing liquid, and ultrasonically processes it to obtain a sample nucleic acid solution.

[0004] For another example, patent application CN118234843A describes an automated nucleic acid extraction box and an automated nucleic acid extraction system including the box. The box has a housing, which includes a sample port, a cell processing chamber, a washing liquid chamber, a filter assembly including a filter element, and a diverter valve having a first reversible sealed output and a second reversible sealed output, wherein the sample port and the cell processing chamber, the cell processing chamber and the filter assembly, and the washing liquid chamber and the filter assembly communicate through a one-way fluid channel, the filter assembly communicates with the diverter valve fluid, and (i) communicates with the waste liquid pipeline when the diverter valve is biased to the first reversible sealed output, and (ii) communicates with the pathogen nucleic acid pipeline when the diverter valve is biased to the second reversible sealed output. However, the structure of this automated nucleic acid extraction system is very complex, and the processing difficulty and application cost of the system are very high, which is not conducive to application scenarios such as large-scale nucleic acid extraction.

[0005] For another example, patent application CN200980133630.3 discloses a disposable device for automated biological sample preparation, which has: a connection mechanism for connecting to a closed ultrasonic treatment tube, and a filter membrane capture unit for capturing samples and processing the samples without the need for other tubes. The operation process of this device is also very cumbersome, such as the need to remove the plug spacer after the ultrasonic treatment tube is placed in an ultrasonic crusher in an automated operation. And the disposable device also needs to be combined with external equipment such as a buffer bottle, a waste bottle, a washing bottle, an elution bottle, and a PCR reagent bottle to complete the extraction of nucleic acids. Among them, other types of buffers and reagents can be used according to the target substance to be separated. The cylinder is preloaded with a measured sample preparation reagent for a single sample preparation operation. The bottle can be sealed with a film at the top to prevent liquid evaporation during storage.

[0006] It can be seen that the above-mentioned automated and manual extraction equipment have the problem of cumbersome operation. With the development of clinical molecular diagnosis, the extraction of pathogen nucleic acid and identification through PCR amplification can be used as a clinical reference, and molecular biology technology also plays an important role in disease diagnosis. Among them, the quality of nucleic acid samples is the key to affecting subsequent results. When the quality of nucleic acid cannot meet the requirements, it will affect the accuracy and detection limit of subsequent test results. In addition, the sample collection and transportation process, extraction method, and efficiency will have an impact on nucleic acid. In addition, general nucleic acid extraction has relatively high requirements for instruments and personnel, and the entire extraction process takes a long time.

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

[0008] The object of the present invention is to provide a nucleic acid extraction device and method, which can partially solve or alleviate the above-mentioned deficiencies in the prior art and simplify the nucleic acid extraction operation.

[0009] In order to solve the above-mentioned technical problems, the present invention specifically adopts the following technical solutions: A nucleic acid extraction device, comprising: A sample adding part, wherein a first accommodating cavity is provided in the sample adding part, and the first accommodating cavity is used to accommodate a first sample liquid; a first end of the sample adding part has an openable cover; A connecting portion, the connecting portion being connected to the sample adding portion, a puncture portion being formed at a first end of the connecting portion, the puncture portion having a puncture end capable of puncturing the wall of the first accommodating cavity, and a first connecting cavity being formed below the puncture portion; The connecting portion has a first surface, a limiting protrusion is arranged on the first surface, the sample adding portion has a second surface, a limiting path is arranged on the second surface, and the limiting path includes: a first limiting segment and a second limiting segment arranged along a direction from the second end to the first end of the sample adding portion, and the first limiting segment and the second limiting segment are arranged to intersect; When the sample adding part and the connecting part are in the first connection state, the limiting protrusion is inserted into the first limiting section, and there is a distance between the puncture end and the first accommodating cavity. When the limiting protrusion moves from the first limiting section into the second limiting section, the sample adding part and the connecting part are converted from the first connection state to the second connection state, and the puncture end punctures the wall of the first accommodating cavity to guide the first sample liquid into the first connecting cavity from the side of the puncture end. A spring, when the sample adding portion is connected to the connecting portion and when the limiting protrusion is inserted into the first limiting section, two ends of the spring are respectively in contact with the end surfaces of the connecting portion and the sample adding portion that are arranged opposite to each other; The reaction part is connected to the connecting part, and the reaction part has a second accommodating cavity therein, and the second accommodating cavity is used to accommodate a second sample liquid.

[0010] 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 arranged to intersect with the second limiting segment; correspondingly, when the sample adding portion and the connecting portion are in the second connection state, a gap is formed between the puncture end and the opening of the sample adding portion; When the limiting protrusion moves from the second limiting section to the third limiting section, the sample adding portion and the connecting portion are converted from the second connection state to the third connection state, and the distance between the puncture end and the opening is reduced so that the puncture end is adjacent to the opening.

[0011] In some embodiments, a fourth limiting segment connected to the third limiting segment and having a certain length is further provided along the direction from the second end to the first end of the connecting portion, and the third limiting segment and the fourth limiting segment are also cross-disposed.

[0012] In some embodiments, the diameter of the puncture portion gradually increases in a direction away from the puncture end, and a plurality of drainage channels are arranged at intervals on the side of the puncture portion to guide the first sample liquid from the puncture end into the first connecting cavity.

[0013] In some embodiments, the puncture portion and the first wall of the connecting portion are connected by a curved second wall, and the second wall is recessed toward a 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, and the drainage surface can limit the flow of liquid to the connection between the sample addition portion and the connecting portion.

[0014] In some embodiments, the first sample liquid stored in the first containing chamber is an acidic liquid.

[0015] In some embodiments, the second sample liquid stored in the reaction part is a lysate.

[0016] In some embodiments, the second end of the reaction portion is inwardly recessed to form a recessed area, and a solid substance for lubrication is smeared on the recessed area.

[0017] The present invention also provides a method for extracting nucleic acid, comprising the steps of: S101 provides a nucleic acid extraction device as described in any one of the embodiments; S102: adding a third sample to be tested into the reaction part, wherein a second sample liquid is also provided in the reaction part; S103 ultrasonically treat the reaction part by using an ultrasonic device, so that the second sample liquid promotes the third sample to release nucleic acid under the action of ultrasound; S104 converts the sample adding part and the connecting part from the first connecting state to the second connecting state, so that the first sample liquid 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.

[0018] In some embodiments, the steps include: S105: converting the sample adding part and the connecting part from the second connecting state to the third connecting state, so that the puncture end is close to the opening of the sample adding part; S106: Invert the nucleic acid extraction device, and the liquid in the reaction part flows out through the opening under the guidance of the puncture end.

[0019] Beneficial technical effects: The present invention provides a nucleic acid extraction device with integrated multifunctional storage and processing of samples. The sample adding part and the connecting part in the extraction device cooperate with the spring through a multi-level limit locking function, and can provide multiple stable connection states (such as a first connection state and a second connection state) to adapt to the operation requirements at different stages.

[0020] Specifically, the cross-setting of the multi-stage limiting segments can assist the user to quickly puncture the first containing cavity to promote the release of the first sample liquid by rotating the connecting portion or the sample adding portion. At the same time, this multi-stage limiting design can also maintain the stability of the device during the transportation of the nucleic acid extraction device. For example, when the nucleic acid extraction device is subjected to bumps, collisions, etc. during transportation, the limiting effect of the limiting segment can prevent the bumps from causing relative rotation between the connecting portion and the sample adding portion, resulting in premature release of the first sample liquid.

[0021] When the user rotates the connection part (or the sample adding part), the spring matched with the multi-stage limit section can provide a certain degree of feedback force to the user. Specifically, when the connection part rotates relative to the sample adding part in the first direction (i.e., the unlocking direction), the distance between the two end faces gradually decreases, the spring contracts, and the force applied by the spring to the end face (equivalent to the feedback force) gradually increases. Therefore, the user can comprehensively judge the rotation operation state at this time through the force perceived by the hand.

[0022] At the same time, the spring can also play a resetting role. Specifically, when the user applies an external force to the connecting part to rotate the connecting part, the puncture end also punctures the first accommodating cavity (the spring is compressed at this time), and when the user cancels the applied external force, the spring can automatically extend under the resetting effect, so that the limiting protrusion returns to the initial position under the resetting effect of the spring.

[0023] The nucleic acid extraction device of the present invention only needs to cooperate with ultrasonic equipment to complete the on-site nucleic acid release, which is conducive to the rapid on-site operation of the detection personnel, and the restriction of the equipment to the site is relatively small. The extraction device of the present invention can be widely used in large-scale detection scenarios such as on-site sampling and off-site detection. It is worth noting that in these large-scale detection scenarios, the number of objects is large, and the sampling and detection work at this time is extremely heavy, and the efficiency requirements are also extremely high. And this large-scale detection activity often involves long-term on-site sampling, off-site detection and other problems. BRIEF DESCRIPTION OF THE DRAWINGS

[0024] In order to more clearly illustrate the technical solutions in the embodiments of the present invention or the prior art, the drawings required for use in the embodiments or the prior art descriptions are briefly introduced below. In all the drawings, similar elements or parts are generally identified by similar reference numerals. In the drawings, each element or part is not necessarily drawn according to the actual scale. Obviously, the drawings described below are some embodiments of the present invention. For those of ordinary skill in the art, other drawings can also be obtained based on these drawings without paying creative labor.

[0025] Figure 1 is a schematic cross-sectional structural diagram of a device in an exemplary embodiment of the present invention; Figure 2 is a schematic structural diagram of a device in an exemplary embodiment of the present invention; Figure 3 It is a schematic diagram of the exploded parts of a device in an exemplary embodiment of the present invention; Figure 4 is a schematic structural diagram of a connecting portion of a device in an exemplary embodiment of the present invention; Figure 5 It is a schematic structural diagram of a sample adding part of a device in an exemplary embodiment of the present invention; Figure 6 It is a schematic structural diagram of a limiting path of a device in an exemplary embodiment of the present invention; Figure 7 It is a schematic structural diagram of a device in another exemplary embodiment of the present invention.

[0026] Summary of reference numerals: sample adding part 10, first accommodating chamber 11, cover body 12, second surface 13, limiting path 14, first limiting section 141, second limiting section 142, third limiting section 143, fourth limiting section 144, first space 15, second space 16; connecting part 20, puncture part 21, drainage channel 211, first connecting chamber 22, first surface 23, limiting protrusion 24, second connecting chamber 25; reaction part 30, second accommodating chamber 31, recessed area 32, spring 40. DETAILED DESCRIPTION

[0027] In order to make the purpose, technical solution and advantages of the embodiments of the present invention clearer, the technical solution in the embodiments of the present invention will be clearly and completely described below in conjunction with the drawings in the embodiments of the present invention. Obviously, the described embodiments are part of the embodiments of the present invention, not all of the embodiments. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative work are within the scope of protection of the present invention.

[0028] Herein, suffixes such as "module", "component" or "unit" used to represent elements are only used to facilitate the description of the present invention, and have no specific meanings by themselves. Therefore, "module", "component" or "unit" can be used mixedly.

[0029] In this document, the terms "upper", "lower", "inner", "outer", "front", "back", "one end", "the other end" and the like indicate positions or positional relationships based on the positions or positional relationships shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the devices or elements referred to must have a specific orientation, be constructed and operate in a specific orientation, and therefore cannot be understood as limiting the present invention. In addition, the terms "first" and "second" are used for descriptive purposes only and cannot be understood as indicating or implying relative importance.

[0030] In this document, unless otherwise clearly specified and limited, the terms "installed", "provided with", "connected", etc. should be understood in a broad sense. 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, or it can be the internal communication of two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0031] Herein "and / or" includes any and all combinations of one or more of the associated listed items.

[0032] Herein, "plurality" means two or more than two, ie, it includes two, three, four, five, etc.

[0033] As used in this specification, the term "about" typically means + / - 5% of the stated value, more typically + / - 4% of the stated value, more typically + / - 3% of the stated value, more typically + / - 2% of the stated value, even more typically + / - 1% of the stated value, and even more typically + / - 0.5% of the stated value.

[0034] In this specification, some embodiments may be disclosed in a format of being in a range. It should be understood that this description of "being in a range" is only for convenience and brevity, and should not be interpreted as a rigid limitation on the disclosed range. Therefore, the description of the range should be considered to have specifically disclosed all possible sub-ranges and independent numerical values ​​within this range. For example, the description of the range 1-6 should be considered to have specifically disclosed sub-ranges 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., as well as individual numbers within this range, such as 1, 2, 3, 4, 5 and 6. Regardless of the breadth of the range, the above rules apply.

[0035] The term "patient" or "subject" is used in this specification to describe an animal, such as a mammal, preferably a human or a domestic animal. In any aspect or specific embodiment described in the present disclosure, the subject or patient may also be a primate. In any aspect or specific embodiment described in the present disclosure, the patient or subject is a human. In any aspect or specific embodiment described in the present disclosure, the patient or subject is livestock, such as cattle, sheep, goats, cows, pigs, etc.; or domestic animals, such as dogs, fish, guinea pigs and cats. In any aspect or specific embodiment described in the present disclosure, the subject is a rodent (e.g., mouse, rat, hamster), rabbit, primate, non-human primate or pig, such as purebred pig, etc.

[0036] In this article, radial refers to the direction passing through the shaft centerline (or axis) in the radial plane. The radial plane refers to the plane perpendicular (or approximately perpendicular) to the shaft centerline.

[0037] In this article, see Figure 6 As shown, when the connecting portion continuously rotates along the first direction, the limiting protrusion continuously moves in the limiting path, and the two ends of the limiting segment along the moving direction can be respectively referred to as the first end and the second end of the limiting segment. The first direction can also be referred to as the unlocking direction, and when the connecting portion moves along the first direction, the distance between the puncture end of the connecting portion and the opening of the sample adding portion gradually decreases. Correspondingly, when the connecting portion continuously rotates along the second direction (opposite to the first direction), the distance between the puncture end of the connecting portion and the opening of the sample adding portion will also gradually increase.

[0038] See also Figure 1-Figure 7 As shown, the present invention provides a nucleic acid extraction device.

[0039] Preferably, the nucleic acid extraction device of the present invention comprises: A sample adding part 10, wherein a first accommodating chamber 11 is provided in the sample adding part 10, and the first accommodating chamber is used to accommodate a first sample liquid (for example, HCl); A connecting portion 20, the connecting portion 20 is connected to the sample adding portion 10, a puncture portion 21 is formed at a first end of the connecting portion, the puncture portion has a puncture end capable of puncturing the wall of the first accommodating cavity, and a first connecting cavity 22 is formed below the puncture portion (i.e., on a side away from the puncture end); The connecting portion 20 has a first surface 23, on which a limiting protrusion 24 is arranged, and the sample loading portion 10 has a second surface 13, on which a limiting path 14 is arranged, and the limiting path 14 includes: a first limiting segment 141 with a certain length and a second limiting segment 142 with a certain length, which are sequentially arranged along the direction from the second end to the first end of the sample loading portion, and the first limiting segment and the second limiting segment 142 are arranged to intersect; When the sample adding part and the connecting part are in the first connection state, the limiting protrusion is stuck in the first limiting section. At this time, there is a certain distance between the puncture end and the first accommodating cavity (specifically, the lower surface of the first accommodating cavity). When the limiting protrusion moves from the first limiting section into the second limiting section, the sample adding part and the connecting part are converted from the first connection state to the second connection state. At this time, the distance between the puncture end and the first accommodating cavity gradually decreases, and the puncture end punctures the wall of the first accommodating cavity to guide the first sample liquid into the first connecting cavity 22 from the side of the puncture end; The reaction part 30 is connected to the connection part, and the reaction part has a second accommodating chamber 31 therein, and the second accommodating chamber is used to accommodate a second sample liquid (eg, NaOH). Specifically, the first connection chamber 22 is connected to the second accommodating chamber 31 .

[0040] In some embodiments, when the released nucleic acid needs to be stored for a long time, the first end of the sample adding part has an openable cover 12, and when the cover is closed, it can play a role in sealing and protecting the nucleic acid.

[0041] Of course, in some on-site sampling and on-site testing scenarios, the sample adding portion only needs to have an opening through which the liquid can flow out.

[0042] Preferably, see Figure 6 As shown, the connected first limiting section 141 and the second limiting section 142 are arranged to intersect each other, so that when the limiting protrusion 24 is inserted into the first limiting section 141, the wall of the first limiting section 141 (which has a certain length) can limit the limiting protrusion 24, thereby preventing the limiting protrusion 24 from being improperly displaced and affecting the stability of the internal liquid; For example, the cross setting of the limiting segments can assist the user to quickly puncture the first containing cavity to promote the release of the first sample liquid by rotating the connecting portion or the sample adding portion. At the same time, this multi-stage limiting design can also maintain the stability of the device during the transportation of the nucleic acid extraction device. For example, when the nucleic acid extraction device is subjected to bumps, collisions, etc. during transportation, the limiting effect of the limiting segments can prevent the bumps from causing relative rotation between the connecting portion and the sample adding portion, resulting in premature release of the first sample liquid.

[0043] Furthermore, the second limiting section can be arranged in a direction parallel or approximately parallel to the radial plane, whereby, when the limiting protrusion is stuck in the middle position of the second limiting section having a certain length, the second limiting section can maintain the puncture section and the first accommodating chamber in a specific connection state, so as to enable the liquid to smoothly enter the first connecting chamber under the guidance of the puncture end, and then enter the reaction part.

[0044] For example, in some embodiments, the limiting path may be a gap that runs through the wall of the sample adding portion, so as to limit the limiting protrusion.

[0045] In some embodiments, it also includes: a spring 40. Correspondingly, when the sample adding part is connected to the connecting part and when the limiting protrusion is inserted into the first limiting section, the two ends of the spring are respectively in contact with the end surfaces of the connecting part and the sample adding part that are arranged opposite to each other.

[0046] Preferably, see Figure 1 As shown, the wall of the sample adding part 10 is formed with a protruding end surface along its radial direction (or approximately radial direction), and the wall of the connecting part 20 is also formed with a protruding end surface along its radial direction (or approximately radial direction), and the two end surfaces are arranged opposite to each other, and a spring 40 is arranged between the two end surfaces. The spring is sleeved on the second surface of the sample adding part and the first surface of the connecting part.

[0047] In this embodiment, when the user rotates the connection part (or the sample adding part), the spring can provide a certain degree of feedback force to the user. Specifically, when the connection part rotates relative to the sample adding part in the first direction (i.e., the direction for puncturing the first accommodating cavity, also referred to as the unlocking direction), the distance between the two end faces gradually decreases, and the spring contracts at this time, and the force (equivalent to the feedback force) applied by the spring to the end face also gradually increases. Therefore, the user can comprehensively judge the rotation operation state at this time through the force perceived by the hand.

[0048] Furthermore, the spring can also play a resetting role. Specifically, when the user applies an external force to the connection part to rotate the connection part so that the limiting protrusion 24 moves from the first end of the first limiting section 141 to the second end, the puncture end also punctures the first accommodating cavity (the spring is compressed at this time); correspondingly, when the user cancels the applied external force, the spring can automatically extend under the resetting effect, so that the limiting protrusion 24 returns to the second end of the first limiting section 141 under the resetting effect of the spring.

[0049] In some embodiments, see Figure 6 As shown, a third limiting section 143 connected to the second limiting section is further provided along the direction from the second end to the first end of the connecting portion, and the third limiting section is arranged to intersect with the second limiting section.

[0050] Correspondingly, when the sample adding part and the connecting part are in the second connection state, a distance is formed between the puncture end and the opening of the sample adding part (this can also avoid excessive liquid accumulation in the opening area and avoid liquid leakage at the opening to a certain extent); when the limiting protrusion moves from the second limiting section to the third limiting section, the sample adding part and the connecting part are converted from the second connection state to the third connection state, and the distance between the puncture end and the opening is reduced so that the puncture end is adjacent to the opening.

[0051] In some embodiments, the diameter of the puncture portion gradually increases in a direction away from the puncture end (in other words, the diameter of the puncture portion gradually increases in a direction from its first end to the second end), and a plurality of drainage channels 211 are arranged at intervals on the side of the puncture portion to guide the first sample liquid from the puncture end into the first connecting cavity.

[0052] In some embodiments, see Figure 7 As shown, the puncture portion 21 is connected to the first wall 25a of the connecting portion 20 (i.e., the wall of the second connecting cavity 25) via a curved second wall 21a, and the second wall 21a is recessed toward a side away from the puncture end; the first wall is tightly fitted to the inner surface of the sample adding portion; For example, see Figure 1As shown, the cavity in the connecting part 20 is divided into a first connecting cavity 22 and a second connecting cavity 25 by the puncture part 21, and the two are respectively used to connect with the reaction part 30 and the sample adding part 10. Specifically, the first wall of the second connecting cavity 25 is formed by extending from the second end of the puncture part 21 in a direction away from the first connecting cavity 22; thus, the first wall, the second wall and the wall surface outside the puncture part 21 cooperate to form a curved drainage surface, and the drainage surface can, on the one hand, guide the liquid to directly enter the first connecting cavity 22 below through the drainage surface during the puncture process of the device (avoiding excessive liquid from entering the boundary between the second space 16 and the connecting part to cause leakage); on the other hand, when the device is inverted to invert the sample, the drainage surface can still play a similar role in preventing leakage.

[0053] Further, in some embodiments, see Figure 6 As shown, along the direction from the second end to the first end of the connecting portion (as shown Figure 6 A fourth limiting segment 144 connected to the third limiting segment 143 and having a certain length is also provided on the upper surface (shown in the middle direction F), and the fourth limiting segment 144 connected to the third limiting segment 143 is also cross-disposed. Preferably, the fourth limiting segment 144 can also be arranged in a direction parallel or approximately parallel to the radial plane, so that when the limiting protrusion 24 is inserted into the fourth limiting segment, the device can be kept in a state where the puncture end is adjacent to the opening.

[0054] Furthermore, after the nucleic acid extraction device has completed the extraction of nucleic acid, the device can be inverted so that the liquid in the device can be released through the opening of the sample adding part (ie, inverted).

[0055] In this embodiment, when the nucleic acid extraction device is inverted, the design of the multi-stage limiting segment can assist the user in operating so that the puncture end is adjacent to the opening of the sample loading part (for example, the puncture end can even be flush with the opening or protrude from the opening). As a result, under the drainage effect of the puncture end, the liquid can flow out directly without flowing too much to the inner surface of the opening (thereby avoiding leakage between the connecting part and the sample loading part to a certain extent). Therefore, when the collected nucleic acid involves different detection items and multiple inversions are required, the multi-stage locking scheme combined with the inverted puncture end can reduce the contamination that may be caused during the inversion process.

[0056] In this embodiment, a multi-stage locking scheme is formed by the relative arrangement of multiple limit segments in direction. This multi-stage locking scheme provides multiple connection states for the nucleic acid extraction device. This multiple connection state can meet the needs of users in different stages such as puncture and sampling, and is simple to operate, which can be achieved by only continuously rotating the connection part.

[0057] In other words, the present invention realizes multi-functional integration (that is, provides an integrated device) through the multi-level locking scheme between the connecting part and the sample adding part. For example, multiple functions such as sample liquid prediction, nucleic acid release, lysis solution synthesis, nucleic acid storage, etc. can be realized through the nucleic acid extraction device.

[0058] Of course, in other embodiments, the limiting path may also be set as a linear path. In this case, the connection state between the connecting portion and the opening portion may be manually controlled by the user.

[0059] Further, in some embodiments, see Figure 4 As shown, the sample adding part 10 is formed with a first space 15 and a second space 16 at both ends of the first accommodating cavity 11. The first space is formed with an opening at one end away from the first accommodating cavity, and the second space 16 is used to accommodate the puncture part.

[0060] Specifically, see Figure 1 As shown, when the sample adding portion and the connecting portion are in a connected state, the inner surface of the sample adding portion (ie, the third surface) and the outer surface of the connecting portion (ie, the second surface) are tightly fitted.

[0061] In some embodiments, a sealing layer (also referred to as an anti-slip layer) may be provided on the inner surface of the sample adding portion and / or the outer surface of the connecting portion. 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, so as to further improve the sealing performance between the sample adding portion and the connecting portion.

[0062] In some embodiments, the first sample liquid is an acidic liquid.

[0063] In some embodiments, the second sample liquid is a lysate. Specifically, the lysate can be an alkaline liquid.

[0064] In some embodiments, the method further comprises: an ultrasonic device having a mounting position for accommodating the reaction part. When the extraction device is arranged in the ultrasonic device, the spring arranged therein can prevent the ultrasonic effect from interfering with the connection state between the connection part and the sample adding part, thereby causing the first sample liquid to be released prematurely.

[0065] In some embodiments, the second end of the reaction portion 30 is recessed inward to form a recessed area 32 , and a solid substance for lubrication is applied on the recessed area 32 .

[0066] Correspondingly, the setting of solid materials can reduce the loss of ultrasonic waves during propagation to a certain extent.

[0067] Furthermore, in some embodiments, a raised area corresponding to the recessed area is formed on the installation position of the ultrasonic device, so that efficient transmission of ultrasonic waves can be achieved through the cooperation between the recessed area and the raised area, and the reaction part can also be fixed.

[0068] In some embodiments, the first end of the reaction portion 30 is rotatably connected to the second end of the connection portion 20 by means of a thread.

[0069] In some embodiments, the sample adding portion is connected to the cover body via threads.

[0070] Alternatively, in some embodiments, the cover body may be a rubber cover with elasticity, and the rubber cover can be inserted into the opening above the sample adding portion to seal the opening.

[0071] In some embodiments, the upper surface and the lower surface of the first accommodating cavity 11 are both formed by a sealing film (such as a film, an aluminum foil film, etc.) to facilitate puncture by the puncture end.

[0072] In some embodiments, the connection part can be directly removed to pour out the liquid in the reaction part, and therefore, the sample adding part may not be provided with an opening for pouring the sample.

[0073] In order to more clearly illustrate the technical solution and beneficial technical effects adopted by the nucleic acid extraction device of the present invention, the extraction method adopted will be mainly explained below by taking whole blood (equivalent to the third sample), HCl (equivalent to the first sample liquid), and NaOH (equivalent to the second sample liquid) as examples: The present invention also provides a method for extracting nucleic acid, comprising the steps of: S101 uses the nucleic acid extraction device described in any one of the embodiments herein; S102: adding a third sample to be tested (such as whole blood) into the reaction part, wherein a second sample liquid is also provided in the reaction part; For example, the sample adding part, the connection part, and the reaction part of the nucleic acid extraction device can be connected in sequence during transportation. When performing the nucleic acid extraction operation, the connection part 20 is first removed to add the sample to be tested (equivalent to the third sample, such as whole blood, serum, swab, feces, urine, sputum, and other different types of samples) through the opening at the first end of the reaction part 30; S103: ultrasonically treating the reaction part by ultrasonic equipment, so that the second sample liquid (such as NaOH) causes the third sample to release nucleic acid under the action of ultrasound; In this embodiment, low-frequency ultrasound is used for treatment, the purpose of which is to generate vibrations to release nucleic acids while maintaining the integrity of nucleic acids. Under the action of ultrasound treatment, NaOH promotes the rapid release of nucleic acids in whole blood.

[0074] Furthermore, a first unlocking operation (or puncture operation) is performed, for example, step S104 is performed to convert the sample adding part and the connecting part from the first connection state to the 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.

[0075] Furthermore, in some embodiments, the method further comprises the step of performing a second unlocking operation to complete the purpose of reversing the sample; specifically, the method comprises the following steps: S105: converting the sample adding part and the connecting part from the second connecting state to the third connecting state, so that the puncture end is close to the opening of the sample adding part; S106: Invert the nucleic acid extraction device, and the liquid in the reaction part flows out through the opening under the guidance of the puncture end.

[0076] In some embodiments, different ultrasonic conditions may be used for different types of third samples, as exemplified below: (1) Oral / nasopharyngeal swabs, vaginal swabs or other swabs, serum samples: Ultrasonic frequency: 60-100kHz, time 3-30 seconds; (2) Sputum: Ultrasonic frequency: 100kHz~300kHz, time 5-60 seconds; The extraction method of the present invention combines ultrasound and strong alkaline lysis, and the device can be used to directly release nucleic acids in sputum or whole blood. In addition, such an integrated structural design can directly neutralize the lysate through a rotary puncture operation, that is, neutralize it with HCl after lysis, thereby avoiding the influence of NaOH on the PCR reaction (also known as polymerase chain reaction). Therefore, it is only necessary to continuously perform ultrasonic strong alkaline lysis and puncture operations on the nucleic acid extraction device to neutralize the lysate, and then use an integrated nucleic acid extraction device to complete the extraction of nucleic acids from samples such as whole blood. In addition, the extracted nucleic acids can be stably stored and transported directly in the device, thereby greatly reducing the labor intensity in the batch nucleic acid extraction process and significantly reducing the workload of the test personnel.

[0077] Furthermore, the nucleic acid extraction device of the present invention can complete on-site nucleic acid release only by cooperating with ultrasonic equipment, which is conducive to the rapid on-site operation of detection personnel, and has relatively few restrictions on the equipment and the site.

[0078] For example, the extraction device of the present invention can be widely used in large-scale detection scenarios such as on-site sampling and off-site detection. It is worth noting that in these large-scale detection scenarios, the number of objects is large, and the sampling and detection work at this time is extremely heavy, and the efficiency requirements are also extremely high. In addition, such large-scale detection activities often involve long-term on-site sampling, off-site detection and other issues.

[0079] For example, it is necessary to collect samples from subjects (such as residents of a community) in an area far from a hospital or testing laboratory for a long time, and then transport the samples to the hospital or testing laboratory. For another example, the subjects may be livestock in a breeding farm in a remote area.

[0080] Since it takes a certain amount of time to transport samples from the testing site to the laboratory, various problems may also occur during this process, such as damage to samples due to improper storage, loss of samples resulting in a decrease in quantity, contamination of liquids during transportation, etc. At the same time, in large-scale testing scenarios, the sampling tasks of testers are very heavy, and the efficiency of nucleic acid sampling and extraction on site is also low.

[0081] In this regard, the present invention proposes an integrated nucleic acid extraction device that can quickly perform operations such as ultrasound, strong alkaline lysis, and neutralization of lysis solution. This nucleic acid extraction device only needs to be configured with an ultrasonic device to complete the extraction of nucleic acids. Therefore, it can effectively reduce the detection cost from the equipment configuration, and at the same time, it can also reduce the manual operation of the detection personnel.

[0082] In addition, the nucleic acid extraction device can stably store the predicted sample liquid in the device through a multi-level locking design. At the same time, the multi-level locking design allows the tester to manually complete operations such as neutralizing the lysis solution and pouring out the nucleic acid by simply turning the connecting part, which greatly reduces the workload of the tester for on-site nucleic acid extraction.

[0083] Furthermore, the multi-level locking design is also beneficial for maintaining the relative stability of the internal structure of the extraction device during long-distance transportation, so as to ensure that it can stably store nucleic acids.

[0084] The continuous ultrasound and strong alkaline lysis operation method implemented by the nucleic acid extraction device has strong versatility and can be applied to different types of samples such as whole blood, serum, swabs, feces, urine, sputum, etc.

[0085] It should be noted that, in this article, the terms "include", "comprises" or any other variations thereof are intended to cover non-exclusive inclusion, so that a process, method, article or device including a series of elements includes not only those elements, but also other elements not explicitly listed, or also includes elements inherent to such process, method, article or device. In the absence of further restrictions, an element defined by the sentence "comprises a ..." does not exclude the existence of other identical elements in the process, method, article or device including the element.

[0086] Through the description of the above implementation methods, those skilled in the art can clearly understand that the above-mentioned embodiment methods can be implemented by means of software plus a necessary general hardware platform, and of course by hardware, but in many cases the former is a better implementation method. Based on such an understanding, the technical solution of the present invention, in essence, or the part that contributes to the prior art, can be embodied in the form of a software product, which is stored in a storage medium (such as ROM / RAM, a magnetic disk, or an optical disk), and includes a number of instructions for a computer terminal (which can be a mobile phone, a computer, a server, or a network device, etc.) to execute the methods described in each embodiment of the present invention.

[0087] The embodiments of the present invention are described above in conjunction with the accompanying drawings, but the present invention is not limited to the above-mentioned specific implementation modes, which are merely illustrative rather than restrictive. Under the guidance of the present invention, ordinary technicians in this field can also make many forms without departing from the scope of protection of the present invention and the claims, all of which are within the protection of the present invention.

Claims

1. A nucleic acid extraction device, characterized in that: include: A sample adding portion (10), wherein a first accommodating cavity (11) is provided in the sample adding portion (10), and the first accommodating cavity is used to accommodate a first sample liquid; a first end of the sample adding portion has an openable cover (12); A connecting portion (20), the connecting portion (20) being connected to the sample adding portion (10), a puncture portion (21) being formed at a first end of the connecting portion, the puncture portion having a puncture end capable of puncturing a wall of the first accommodating cavity, and a first connecting cavity (22) being formed below the puncture portion; The connecting portion (20) has a first surface (23) on which a limiting protrusion (24) is arranged, and the sample loading portion (10) has a second surface (13) on which a limiting path (14) is arranged, and the limiting path (14) comprises: a first limiting segment (141) and a second limiting segment (142) arranged in a direction from the second end to the first end of the sample loading portion, and the first limiting segment and the second limiting segment (142) are arranged to intersect; When the sample adding part and the connecting part are in the first connection state, the limiting protrusion is inserted into the first limiting section, and there is a distance between the puncture end and the first accommodating cavity. When the limiting protrusion moves from the first limiting section into the second limiting section, the sample adding part and the connecting part are converted from the first connection state to the second connection state, and the puncture end punctures the wall of the first accommodating cavity to guide the first sample liquid into the first connecting cavity from the side of the puncture end. a spring (40), wherein when the sample adding portion is connected to the connecting portion and when the limiting protrusion is inserted into the first limiting section, two ends of the spring are respectively in contact with end surfaces of the connecting portion and the sample adding portion that are arranged opposite to each other; A reaction part (30), the reaction part (30) being connected to the connection part, the reaction part having a second accommodating cavity (31) therein, the second accommodating cavity being used to accommodate a second sample liquid.

2. A nucleic acid extraction device according to claim 1, characterized in that: A third limiting section (143) connected to the second limiting section is also provided along the direction from the second end to the first end of the connecting portion, and the third limiting section and the second limiting section are arranged to intersect each other; correspondingly, when the sample adding portion and the connecting portion are in the second connection state, a spacing is formed between the puncture end and the opening of the sample adding portion; When the limiting protrusion moves from the second limiting section to the third limiting section, the sample adding portion and the connecting portion are converted from the second connection state to the third connection state, and the distance between the puncture end and the opening is reduced so that the puncture end is adjacent to the opening.

3. A nucleic acid extraction device according to claim 2, characterized in that: A fourth limiting section (144) connected to the third limiting section (143) and having a certain length is also provided along the direction from the second end to the first end of the connecting portion, and the third limiting section (143) and the fourth limiting section (144) are arranged crosswise.

4. A nucleic acid extraction device according to claim 1, characterized in that: The diameter of the puncture portion gradually increases in a direction away from the puncture end, and a plurality of drainage channels are arranged at intervals on the side surface 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 claim 4, characterized in that: The puncture portion and the first wall of the connecting portion are connected via a curved second wall, and the second wall is recessed toward a 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, and the drainage surface can limit the flow of liquid toward the connection between the sample addition portion and the connecting portion.

6. A nucleic acid extraction device according to claim 1, characterized in that: The first sample liquid stored in the first containing chamber is an acidic liquid.

7. A nucleic acid extraction device according to claim 1, characterized in that: The second sample liquid stored in the reaction part is a lysate.

8. A nucleic acid extraction device according to claim 1, characterized in that: The second end of the reaction part (30) is inwardly recessed to form a recessed area, and a solid substance for lubrication is smeared on the recessed area.

9. A method for extracting nucleic acid, characterized in that: Includes steps: S101 provides a nucleic acid extraction device as described in any one of claims 1 to 8; S102: adding a third sample to be tested into the reaction part, wherein a second sample liquid is also provided in the reaction part; S103 ultrasonically treat the reaction part by using an ultrasonic device, so that the second sample liquid promotes the third sample to release nucleic acid under the action of ultrasound; S104 converts the sample adding part and the connecting part from the first connecting state to the second connecting state, so that the first sample liquid in the connecting part enters the first connecting cavity through the side of the puncture end, and the first sample liquid and the second liquid sample undergo a neutralization reaction.

10. A method for extracting nucleic acid according to claim 9, characterized in that: Also includes the steps: S105: converting the sample adding part and the connecting part from the second connecting state to the third connecting state, so that the puncture end is close to the opening of the sample adding part; S106: Invert the nucleic acid extraction device, and the liquid in the reaction part flows out through the opening under the guidance of the puncture end.

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