CN216237022U

CN216237022U - Nucleic acid detection device

Info

Publication number: CN216237022U
Application number: CN202122304246.6U
Authority: CN
Inventors: 杨医嘉; 解亚平; 戴立忠
Original assignee: Sansure Biotech Inc
Current assignee: Sansure Biotech Inc
Priority date: 2021-09-23
Filing date: 2021-09-23
Publication date: 2022-04-08
Anticipated expiration: 2031-09-23

Abstract

The invention provides a nucleic acid detection device, which comprises a cracking piece, a liquid transfer piece, an amplification piece, a conducting piece and a limiting piece, wherein the cracking piece is internally provided with a cracking cavity and a first through hole communicated with the cracking cavity; the limiting piece is arranged on the inner wall of the liquid transferring piece and limits the displacement of the conducting piece in the first direction. The nucleic acid detection device provided by the invention changes the distance from the lower end of the conducting piece to the bottom end of the liquid transferring cavity by controlling the conducting piece to move in the liquid transferring cavity, and realizes quantitative liquid transferring by limiting the maximum moving displacement of the conducting piece through the limiting piece.

Description

Nucleic acid detection device

Technical Field

The invention relates to the technical field of biology, in particular to a nucleic acid detection device.

Background

Nucleic acid detection is an important detection method for virus detection, and viruses are detected by extracting nucleic acid, amplifying the nucleic acid in a closed space, and reacting the amplified nucleic acid with a detection reagent. The loop-mediated isothermal amplification (LAMP) technology is to design 4-6 specific primers aiming at 6-8 regions of a target gene by using strand displacement DNA polymerase, and to realize 10 specific primers within dozens of minutes under the isothermal condition⁹-10¹⁰Techniques for secondary amplification. The loop-mediated isothermal amplification (LAMP) has the characteristics of high speed, simplicity, high sensitivity and specificity, has low requirements on instruments and experimental sites, and is widely applied to multiple fields of scientific research and nucleic acid detection. The related nucleic acid detection device is provided with a cracking device, an amplification device and a detection device which are all independently arranged, the cracking device, the amplification device and the detection device need to be frequently opened to carry out adding and taking-out operations in the detection process, the steps are multiple, the operation difficulty is high, the use is inconvenient, a quantitative device needs to be additionally added when the detection liquid is transferred among the devices, the liquid quantitative liquid transfer is realized, and the detection complexity is increased.

Disclosure of Invention

In view of the above, embodiments of the present invention provide a nucleic acid detecting apparatus to solve the technical problem of how to simplify the structure of quantitative pipetting.

In order to achieve the purpose, the technical scheme of the invention is realized as follows:

the embodiment of the invention provides a nucleic acid detection device, which comprises: the cracking piece is internally provided with a cracking cavity, and is provided with a first through hole communicated with the cracking cavity; the liquid transferring piece is internally provided with a liquid transferring cavity, the liquid transferring piece is provided with a second through hole and a third through hole which are communicated with the liquid transferring cavity, and the second through hole is communicated with the first through hole; the amplification piece is internally provided with an amplification cavity, the amplification cavity is provided with a fourth through hole communicated with the amplification cavity, and the fourth through hole is communicated with the third through hole; the conducting piece is at least partially arranged in the pipetting cavity and can rotate around a first direction so as to simultaneously close the second through hole and the third through hole and open one of the second through hole and the third through hole; the conducting piece can reciprocate along a first direction to input and output liquid of the liquid transferring cavity according to a set amount; and the limiting part is arranged on the inner wall of the liquid transferring part so as to limit the displacement of the conducting part in the first direction.

In some embodiments, the conducting member includes a small diameter portion and a first large diameter portion extending in the first direction and connected to each other, and the first large diameter portion is close to the bottom end of the liquid transferring cavity relative to the small diameter portion; one end of the first large diameter part, which is close to the small diameter part, is abutted against the inner wall of the liquid transferring piece, one end of the first large diameter part, which is far away from the small diameter part, is provided with an opening communicated with the liquid transferring cavity, and the opening can be rotatably communicated with the second through hole or the third through hole; a space is provided between the small diameter portion and the inner wall.

In some embodiments, the stopper is a protrusion protruding from the inner wall of the pipetting member, the protrusion is located at a distance from the bottom end of the pipetting cavity greater than the length of the first large diameter portion in the first direction, and the protrusion is spaced from or in contact with the small diameter portion.

In some embodiments, the projections are circumferentially disposed about the inner wall of the pipetting member.

In some embodiments, the opening extends along the first direction, and the length of the opening along the first direction is greater than the distance from the second through hole and the third through hole to the bottom end of the pipetting cavity.

In some embodiments, the distance from the second through hole to the bottom end of the pipetting cavity is the same as the distance from the third through hole to the bottom end of the pipetting cavity.

In some embodiments, a ratio of a cross-sectional area of the opening to a cross-sectional area of the first large diameter portion is less than 1/10.

In some embodiments, the lead through further comprises: the second large-diameter part extends along the first direction and is connected to one end, far away from the first large-diameter part, of the small-diameter part, and the second large-diameter part protrudes out of the liquid moving cavity.

In some embodiments, the cross-sectional area of the second large diameter portion is the same as the cross-sectional area of the first large diameter portion at the end thereof near the small diameter portion.

In some embodiments, the lysis member and the amplification member are located on either side of the pipetting member.

The nucleic acid detection device provided by the embodiment of the invention comprises a cracking piece, a liquid transfer piece, an amplification piece, a conducting piece and a limiting piece, wherein the cracking piece is provided with a cracking cavity and a first through hole communicated with the cracking cavity, the liquid transfer piece is provided with a liquid transfer cavity, a second through hole and a third through hole communicated with the liquid transfer cavity, the amplification piece is provided with an amplification cavity and a fourth through hole communicated with the amplification cavity, the first through hole is communicated with the second through hole, the third through hole is communicated with the fourth through hole, at least part of the conducting piece is positioned in the liquid transfer cavity, and can rotate in the liquid transfer cavity and reciprocate in the first direction. The limiting piece is arranged on the inner wall of the liquid transferring piece to limit the displacement of the conducting piece in the first direction. The nucleic acid detection device provided by the embodiment of the invention can simultaneously seal the second through hole and the third through hole by controlling the conducting piece to rotate in the liquid transferring cavity, open the second through hole to communicate the cracking cavity and the liquid transferring cavity, or open the third through hole to communicate the liquid transferring cavity and the amplification cavity, and simultaneously change the distance between the lower end of the conducting piece and the bottom end of the liquid transferring cavity by moving the conducting piece up and down in the first direction, so that the space for accommodating liquid in the liquid transferring cavity is changed, and the limiting piece limits the maximum displacement to the movement of the conducting piece, so that the maximum space for accommodating liquid in the liquid transferring cavity is limited, and quantitative liquid transferring is realized.

Drawings

FIG. 1 is a sectional view of a nucleic acid detecting apparatus from a perspective according to an embodiment of the present invention;

FIG. 2A is a partial sectional view of a nucleic acid detecting apparatus in a state provided by an embodiment of the present invention;

FIG. 2B is a partial sectional view of the nucleic acid detecting apparatus in another state according to the embodiment of the present invention;

FIG. 2C is a partial sectional view of the nucleic acid detecting apparatus in another state according to the embodiment of the present invention;

FIG. 3 is a cross-sectional view of a nucleic acid detecting apparatus from another perspective according to an embodiment of the present invention;

FIG. 4 is a cross-sectional view of a nucleic acid detecting apparatus from another perspective provided in an embodiment of the present invention;

fig. 5 is a schematic structural diagram of a conducting element according to an embodiment of the present invention;

FIG. 6 is an overall cross-sectional view of a lysis member, a pipetting member and an amplification member according to an embodiment of the invention.

Description of reference numerals:

10. a cracking member; 11. a lysis chamber; 12. a first through hole; 13. a sample inlet; 20. a pipetting member; 21. a pipetting chamber; 22. a second through hole; 23. a third through hole; 30. an amplification element; 31. an amplification chamber; 32. a fourth via hole; 33. a feeding port; 40. a conducting piece; 41. a small diameter part; 42. a first large diameter portion; 421. an opening; 43. a second large diameter portion; 50. and a limiting member.

Detailed Description

In order to make the objects, technical solutions and advantages of the present invention more apparent, the present invention is described in further detail below with reference to the accompanying drawings and embodiments. It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.

Various combinations of the specific features in the embodiments described in the detailed description may be made without contradiction, for example, different embodiments may be formed by different combinations of the specific features, and in order to avoid unnecessary repetition, various possible combinations of the specific features in the present application will not be described separately.

In the following description, the term "first \ second \ … …" is referred to merely to distinguish different objects and does not indicate that there is identity or relationship between the objects. It should be understood that the references to "upper", "lower", "upward", "downward" and "bottom" are all intended to refer to the orientation during normal use.

It should be noted that the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other like elements in a process, method, article, or apparatus that comprises the element. The term "coupled", where not otherwise specified, includes both direct and indirect connections.

The nucleic acid detection device provided by the invention can detect nucleic acid in a short time, and is mainly applied to the scene of on-site instant detection, namely on-site instant detection, which mainly means that rapid detection is carried out on the site outside a laboratory, so that the detection time is shortened, and the detection process is accelerated.

The following is an exemplary illustration of the detection scheme for nucleic acid detection:

in the nucleic acid detection, after a detection sample is obtained by a detector, the detection sample is placed into a cracking container for cracking reaction, cell lysate can be placed in the cracking container, the detection sample is contacted with the cell lysate and reacts, a cell membrane of the detection sample is dissolved, and cell contents are released, wherein the cell contents comprise nucleic acid, protein and other substances. The cell lysate is added with an enzyme for dissolving protein to reduce the protein content of the cells so as to improve the ratio of nucleic acid. And then transferring the solution containing the nucleic acid into an amplification container, amplifying the nucleic acid to increase the content of the nucleic acid, transferring the amplified nucleic acid into a detection container after the amplification of the nucleic acid is finished, and reacting the amplified nucleic acid with a detection reagent to obtain a detection result so as to finish the detection process of the nucleic acid. In the above detection process, quantitative transfer of liquid is required to be achieved from the disposable material to the transfer to the amplification vessel, so as to obtain an accurate detection result.

An embodiment of the present invention provides a nucleic acid detecting apparatus, which is shown in fig. 1 and 3, and includes: lysis unit 10, pipetting unit 20, amplification unit 30, conducting unit 40, and stopper 50.

As shown in fig. 1, a hollow cracking chamber 11 is provided inside the cracking member 10, and the cracking member 10 is provided with a first through hole 12 communicating with the cracking chamber 11. The lysis member 10 is a device for performing a lysis reaction on cells to release the contents of the cells, wherein the lysis chamber 11 is used for accommodating a lysis solution and a nucleic acid sample, and the lysis solution is a surfactant for breaking cell membranes of the cells to release the contents of the cells, thereby exposing the nucleic acids. A portion of the lysis member 10 may surround to form a first through-hole 12, the first through-hole 12 communicating with the lysis chamber 11 such that the liquid containing nucleic acids in the lysis chamber 11 can be transferred out of the lysis chamber 11 through the first through-hole 12. The cracking piece 10 can be further provided with a sample inlet 13, the sample inlet 13 can be arranged at the upper end of the cracking piece 10 to communicate the cracking cavity 11 with the external space, and the nucleic acid sample is put into the cracking cavity 11 through the sample inlet 13. A sealing film can be arranged at the sample inlet 13 to seal the sample inlet 13 so as to separate the cracking cavity 11 from the external space, and the sealing film is attached to the sample inlet 13 after the nucleic acid sample is put into the cracking cavity 11. In some embodiments, a closing cap can be disposed at the sample inlet 13 to open and close the sample inlet 13, and when the nucleic acid sample needs to be dispensed into the lysis member 10, the closing cap can be opened and closed after the nucleic acid sample is dispensed.

In some embodiments, the temperature of the cracking unit 10 can be controlled according to the temperature required by the cracking reaction, for example, a thermostatic heating plate can be attached to the outer wall of the cracking unit 10 to control the temperature in the cracking chamber 11 to a predetermined reaction temperature.

As shown in FIG. 1, the pipetting device 20 has a hollow pipetting chamber 21 therein, the pipetting device 20 is provided with a second through hole 22 and a third through hole 23 communicating with the pipetting chamber 21, and the second through hole 22 communicates with the first through hole 12. The pipetting member 20 is a device for temporarily storing a volume of liquid, wherein the pipetting cavity 21 is intended to receive liquid. The pipetting piece 20 is partially surrounded to form a second through hole 22 and a third through hole 23, the second through hole 22 is not directly communicated with the third through hole 23, the second through hole 22 is communicated with the first through hole 12 to transfer the liquid in the lysis chamber 11 to the pipetting chamber 21, and the third through hole 23 is used for transferring the liquid in the pipetting chamber 21 out of the pipetting chamber 21, namely, the liquid in the lysis chamber 11 is transferred to the pipetting chamber 21 through the first through hole 12 and the second through hole 22 and then is transferred out of the pipetting chamber 21 through the third through hole 23.

As shown in FIG. 1, the amplification product 30 has a hollow amplification chamber 31 therein, the amplification chamber 31 has a fourth through-hole 32 communicating with the amplification chamber 31, and the fourth through-hole 32 communicates with the third through-hole 23. The amplification part 30 is a device for performing an amplification reaction of nucleic acid to increase the content of nucleic acid, wherein the amplification chamber 31 is used for containing a liquid containing nucleic acid. A portion of the amplification member 30 may surround to form a fourth through hole 32, and the fourth through hole 32 communicates with the third through hole 23 to transfer the liquid in the pipetting cavity 21 to the amplification cavity 31. Substances such as primer hexamers, phi 29DNA polymerase, deoxyinosine-like primers and the like for promoting the amplification of nucleic acid are required to be placed in the amplification cavity 31, and when liquid containing the nucleic acid enters the amplification cavity 31 and contacts with the substances, the amplification of the nucleic acid is promoted. The amplification part 30 may be provided with a feed port 33, and the feed port 33 may be provided at the upper end of the amplification part 30 to communicate the amplification chamber 31 with the external space so as to administer a substance for promoting the amplification of nucleic acid. A sealing membrane may be disposed at the feeding port 33 to seal the feeding port 33 and isolate the amplification chamber 31 from the external space. In some embodiments, a closing cover can be disposed at the dispensing opening 33 to open and close the dispensing opening 33, and when a substance for promoting nucleic acid amplification needs to be dispensed into the amplification chamber 31, the closing cover can be opened to connect the amplification chamber 31 with the external space; after the administration of the nucleic acid amplification promoting substance is completed, the closing lid may be closed to block the amplification chamber 31 from the external space.

In some embodiments, a nucleic acid detection reagent may be further disposed in the amplification chamber 31 to detect a nucleic acid, for example, to detect whether the nucleic acid is positive or negative. The nucleic acid detection reagent may be fed into the amplification chamber 31 through the feed port 33 so that the amplification of the nucleic acid can be completed or the detection of the nucleic acid can be performed in one amplification chamber 31.

In some embodiments, the temperature of the amplification part 30 may be controlled according to the temperature required for the amplification reaction, for example, a thermostatic heating plate may be attached to the outer wall of the amplification part 30 to control the temperature in the amplification chamber 31 to a predetermined reaction temperature.

It should be noted that the lysis member 10, the pipetting member 20 and the amplification member 30 may be integrally formed to form an integral device, or may be individually formed to be connected to each other, or may be formed as an integral device by two. For example, a lysis chamber 11, a pipetting chamber 21 and an amplification chamber 31 are formed inside a complete housing, and the pipetting chamber 21 is communicated with the lysis chamber 11 and the amplification chamber 31 respectively, so that the liquid in the lysis chamber 11 can be transferred to the pipetting chamber 21 and then transferred from the pipetting chamber 21 to the amplification chamber 31.

The conducting member 40 is at least partially disposed in the pipetting cavity 21 and is rotatable about a first direction (up and down direction as shown in fig. 1) to simultaneously close the second through hole 22 and the third through hole 23 and open one of the second through hole 22 and the third through hole 23. The part of the conducting member 40 located in the pipetting cavity 21 can rotate instantaneously or anticlockwise along the inner wall of the pipetting member 20 to simultaneously close the second through hole 22 and the third through hole 23 or open the second through hole 22 and the third through hole 23, but only one of the second through hole 22 and the third through hole 23 can be opened to control the liquid input or output to the pipetting cavity 21 by controlling the conducting member 40, so that the liquid in the lysis cavity 11 and the liquid in the amplification cavity 31 can not be polluted. The conduction member 40 is reciprocally movable in a first direction (up and down direction as shown in fig. 1) to input and output the liquid of the pipetting chamber 21 by a set amount. The pipetting chamber 21 may communicate with the external space at the upper end in the first direction (up-down direction as shown in fig. 1) so that a portion of the conducting member 40 may protrude out of the pipetting chamber 21, thereby providing a sufficient space for the conducting member 40 to move up and down in the pipetting chamber 21.

When pipetting is carried out, the conducting piece 40 can be rotated to be communicated with the cavities needing to be communicated, and then the conducting piece 40 is controlled to move upwards or downwards to provide power for transferring liquid between the cavities. The operation of the conducting member 40 for controlling the liquid input or output of the pipetting cavity 21 will be described in detail with reference to fig. 2A, 2B and 2C.

As shown in FIG. 2A, the conducting member 40 projects into the pipetting chamber 21, and the bottom end of the conducting member 40 is in contact with the bottom end of the pipetting chamber 21. When the nucleic acid sample is subjected to a lysis reaction in the lysis chamber 11, the lysis chamber 11 and the liquid transfer chamber 21 should be in a separated state, and the liquid transfer chamber 21 should also be separated from the amplification chamber 31 to prevent a substance in the amplification chamber 31 for promoting nucleic acid amplification from entering the liquid transfer chamber 21, so that the conduction piece 40 blocks the second through hole 22 and the third through hole 23, and the liquid transfer chamber 21 is separated from the lysis chamber 11 and also separated from the amplification chamber 31.

As shown in FIG. 2B, when the lysis reaction in the lysis chamber 11 is completed, the conduction member 40 is rotated to connect the second through hole 22 with the pipetting chamber 21, and at this time, the pipetting chamber 21 only needs to be connected with the lysis chamber 11 and kept isolated from the amplification chamber 31, so that the liquid in the lysis chamber 11 can enter the pipetting chamber 21. The conducting piece 40 keeps communicating the lysis cavity 11 and the pipetting cavity 21, separates the pipetting cavity 21 from the amplification cavity 31, and moves the conducting piece 40 upwards to enable the conducting piece 40 to be far away from the cavity bottom of the pipetting cavity 21, so that the space for containing liquid in the pipetting cavity 21 is increased, the distance from the lower end of the conducting piece 40 to the front of the cavity bottom of the pipetting cavity 21 is controlled, the volume of the liquid contained in the pipetting cavity 21 can be controlled, and the amount of the liquid in the lysis cavity 11 transferred to the pipetting cavity 21 can be controlled, so that the liquid can be quantitatively transferred. The conducting member 40 can be a piston-like structure, when the conducting member 40 moves upwards, the pressure in the pipetting cavity 21 can be reduced, the pressure in the lysis cavity 11 can be higher than the pressure in the pipetting cavity 21, and in order to balance the pressures in the pipetting cavity 21 and the lysis cavity 11, the pipetting cavity 21 can suck the liquid in the lysis cavity 11 through the first through hole 12 and the second through hole 22, so that power can be provided for transferring the liquid from the lysis cavity 11 to the pipetting cavity 21.

As shown in FIG. 2C, when the liquid in the pipetting cavity 21 reaches a predetermined amount, the rotary conduction member 40 closes the lysis cavity 11 and the pipetting cavity 21 and connects the pipetting cavity 21 and the amplification cavity 31, and the liquid in the pipetting cavity 21 can enter the amplification cavity 31 from the pipetting cavity 21. Keeping the liquid transferring cavity 21 isolated from the lysis cavity 11 and communicating with the amplification cavity 31, and moving the conducting piece 40 downwards to make the lower end of the conducting piece 40 close to the bottom end of the liquid transferring cavity 21, so as to press the liquid in the liquid transferring cavity 21, so that the liquid is output from the third through hole 23 to the fourth through hole 32 and is transmitted into the amplification cavity 31, and when the lower end of the conducting piece 40 contacts with the cavity bottom of the liquid transferring cavity 21, the liquid in the liquid transferring cavity 21 can be completely pressed into the amplification cavity 31, so that quantitative liquid transferring is realized.

It should be noted that the lysis member 10, the pipetting member 20, the amplification member 30 and the conduction member 40 can be made of transparent materials, so that the nucleic acid testing personnel can directly observe the internal conditions of the lysis member 10, the pipetting member 20, the amplification member 30 and the conduction member 40, so as to conveniently observe the transfer condition of the liquid in the nucleic acid testing device and the adjustment condition of the conduction member 40.

As shown in fig. 3, a stopper 50 is provided on an inner wall of the pipetting member 20 to restrict the displacement of the conducting member 40 in a first direction (up-down direction as shown in fig. 1). The conducting piece 40 moves up and down in the pipetting cavity 21 to change the liquid containing space of the pipetting cavity 21, and the limiting piece 50 is arranged in the pipetting piece 20 to limit the displacement of the conducting piece 40 moving up and down, so that the volume of the liquid containing space in the pipetting cavity 21 can be determined, the amount of liquid in the pipetting cavity 21 is constant during each input and output, and quantitative pipetting is realized. In some embodiments, the limiting member 50 may be a groove disposed on the inner wall of the pipetting member 20, and the small diameter portion 41 of the conducting member 40 is provided with a corresponding elastic boss, the small diameter portion 41 moves up and down to drive the elastic boss to move up and down, and when the elastic boss is snapped into the groove, the movement of the small diameter portion 41 can be limited, so as to fix the volume of the pipetting cavity 21 for containing liquid. After the elastic lug boss is clamped in the groove, the force for pushing and pulling the conducting piece 40 can be increased, so that the elastic lug boss slides out of the groove to release the limiting fixation.

In some embodiments, as shown in fig. 2A-2C, the conducting member 40 includes a small diameter portion 41 and a first large diameter portion 42 extending along the first direction and connected to each other, and the first large diameter portion 42 is close to the bottom end of the liquid moving chamber 21 relative to the small diameter portion 41. The first large diameter portion 42 is entirely located in the pipetting chamber 21 and can be rotated in the pipetting chamber 21 for simultaneously closing the second through-hole 22 and the third through-hole 23 and opening one of the second through-hole 22 and the third through-hole 23. The small diameter portion 41 is connected to the first large diameter portion 42 to move the first large diameter portion 42 up and down in the liquid transferring chamber 21. One end of the first large diameter portion 42 close to the small diameter portion 41 abuts against the inner wall of the liquid transfer member 20. The edge of the cross section of the first large diameter portion 42 in the first direction (the up-down direction shown in fig. 2C) is in contact with the inner wall of the pipetting member 20 to block the pipetting cavity 21, thereby confining the liquid entering the pipetting cavity 21 in the space between the lower end of the first large diameter portion 42 to the bottom end of the pipetting cavity 21. An opening 421 communicated with the liquid transferring cavity 21 is formed in one end, far away from the small diameter part 41, of the first large diameter part 42, and the opening 421 can be rotatably communicated with the second through hole 22 or the third through hole 23. The opening 421 may extend along a first direction (e.g., up and down direction as shown in fig. 2C), and one end of the opening 421 is communicated with the liquid transferring chamber 21, and the other end of the opening does not penetrate the first large diameter portion 42, so that the opening 421 is communicated with the liquid transferring chamber 21 and the first large diameter portion 42 does not block one end of the liquid transferring chamber 21. Rotation of first large diameter portion 42 causes opening 421 to rotate therewith, so that the position at which opening 421 rotates can be adjusted. The opening 421 is rotated to be opposite to the second through hole 22 to communicate the second through hole 22 with the pipetting cavity 21 so that the liquid in the lysis cavity 11 can be transferred to the pipetting cavity 21. The opening 421 is rotated to be opposite to the third through hole 23 to communicate the third through hole 23 with the pipetting cavity 21 so that the liquid in the pipetting cavity 21 can be transferred into the amplification cavity 31. The small diameter portion 41 has a space with the inner wall. The cross-sectional dimension of the small diameter portion 41 in the first direction (the up-down direction shown in fig. 2C) is smaller than the cross-sectional dimension of the first large diameter portion 42 so that the small diameter portion 41 does not contact the inner wall of the pipetting member 20, and the inner wall of the pipetting member 20 does not affect the up-down movement of the small diameter portion 41 in the pipetting chamber 21.

Through seting up the opening in first major diameter portion department to intercommunication second through-hole or third through-hole, thereby make and move liquid chamber and schizolysis chamber or amplify the chamber intercommunication, realize during liquid shifts the different cavitys, and minor diameter portion is connected with first major diameter portion, drives first major diameter portion and reciprocates in moving liquid chamber, in order to realize the ration and move the liquid.

In some embodiments, as shown in FIG. 3, the stop member 50 is a protrusion protruding from the inner wall of the pipetting member 20, the protrusion is located at a distance from the bottom end of the pipetting cavity 21 greater than the length of the first large diameter portion 42 in the first direction, and the protrusion is spaced from or in contact with the small diameter portion 41. The stopper 50 may be a protrusion facing the small diameter portion 41, and the protrusion may have a certain interval with the small diameter portion 41 so as not to affect the movement of the small diameter portion 41; the projection may contact the small diameter portion 41 without preventing the movement of the small diameter portion 41. The distance from the projection to the bottom end of the pipetting cavity is greater than the distance from the upper end to the lower end of the first large diameter portion 42 so that the first large diameter portion 42 can move between the projection and the bottom end of the pipetting cavity 21. The first large diameter portion 42 contacts with the inner wall of the pipetting member 20, so when the first large diameter portion 42 moves to the projection, it is lifted and stopped, and cannot move upwards, thereby defining the liquid containing space of the pipetting cavity 21 for quantitative pipetting.

In some embodiments, as shown in FIG. 4, the projections are disposed about the circumference of the inner wall of the pipetting member 20. The number of the bulges can be multiple, the bulges are arranged in a surrounding mode, and a space exists between every two adjacent bulges. The distance from the plurality of bulges to the bottom end of the liquid transferring cavity 21 is the same, and the plurality of bulges limit the moving displacement of the first large-diameter part together, so that accurate and effective limiting is realized.

In some embodiments, as shown in fig. 5 and 6 in combination, the opening 421 extends in the first direction, and the length of the opening 421 in the first direction (up-down direction as shown in fig. 5) is greater than the distance from the second through hole 22 and the third through hole 23 to the bottom end of the pipetting cavity 21. The opening 421 is a long opening, one end of which is communicated with the liquid transferring cavity 21, and the other end of which extends upward but does not penetrate through the first large diameter portion 42, so as to ensure that the upper end of the first large diameter portion 42 blocks the upper end of the liquid transferring cavity 21. The first through hole 22 and the second through hole 23 may be circular through holes or other shapes such as square holes. The size of the second through hole 22 is smaller than the length of the opening 421 in the vertical direction shown in fig. 6, so that when the second through hole 22 communicates with the opening 421, the second through hole 22 remains in communication with the opening 421 even if the opening 421 moves vertically. The size of the third through hole 23 is smaller than the length of the opening 421 in the vertical direction shown in fig. 6, so that the third through hole 23 is communicated with the opening 421 even if the opening 421 moves up and down. The distance from the second through hole 22 and the third through hole 23 to the bottom end of the pipetting cavity 21 is smaller than the length of the opening 421 (the length in the vertical direction as shown in fig. 6) so that the bottom end of the first large diameter portion 42 can contact the bottom of the pipetting cavity 21 when the first large diameter portion 42 moves downward, and the liquid in the pipetting cavity 21 can be completely squeezed out.

In some embodiments, as shown in connection with fig. 5 and 6, the second through hole 22 is at the same distance from the bottom end of the pipetting cavity 21 as the third through hole 23 is at the bottom end of the pipetting cavity 21. The positions of the second through hole 22 and the third through hole 23 in the up-down direction shown in fig. 6 are the same as the distance from the bottom end of the pipetting cavity 21, so that when the opening 421 is communicated with the second through hole 22 and the third through hole 23 respectively, the upward distance of the opening 421 is the same as the downward moving distance of the opening 421, and therefore the opening 421 can be communicated with the second through hole 22 and simultaneously block the third through hole 23, or the opening 421 is communicated with the third through hole 23 and simultaneously block the second through hole 22, so that one-side communication is realized.

In some embodiments, as shown in fig. 5, the ratio of the cross-sectional area of opening 421 to the cross-sectional area of first large diameter portion 42 is less than 1/10. The size of the opening 421 is far smaller than that of the first large-diameter portion 42, so that the opening of the opening 421 does not affect the blocking of the first large-diameter portion 42 on the upper end of the liquid transferring cavity 21, and when liquid is transferred, the amount of liquid retained in the opening is relatively small and can be ignored, and further, the quantitative transfer of the liquid is not affected.

In some embodiments, as shown in fig. 5, the conducting element 40 further includes a second large diameter portion 43 extending along the first direction (the up-down direction shown in fig. 5) and connected to an end of the small diameter portion 41 away from the first large diameter portion 42, and the second large diameter portion 43 protrudes from the liquid moving chamber 21. One end of the small diameter portion 41 far away from the first large diameter portion 42 is connected with a second large diameter portion 43, and the second large diameter portion 43 always protrudes out of the liquid transferring cavity 21 when the conducting member 40 moves up and down, so that a detector can conveniently control the movement of the conducting member 40 manually. The second large diameter portion 43 may be a cylinder to facilitate rotation. The cross-sectional area of the second large-diameter portion 43 is the same as the cross-sectional area of the first large-diameter portion 42 at the end near the small-diameter portion 41. The second large diameter portion 43 and the first large diameter portion 42 may be configured as the same cylinder to facilitate the machining process of the conduction member 40.

In some embodiments, as shown in FIG. 6, the lysis member 10 and the amplification member 30 are located on either side of the pipetting member 20. The lysis member 10, the amplification member 30 and the pipetting member 20 may be arranged in a line, wherein the pipetting member 20 is located in the middle, so that the conducting member 40 has a sufficient rotation range in the pipetting cavity 21 to facilitate control of the isolation and communication of the lysis chamber 11, the pipetting cavity 21 and the amplification chamber 31.

The above description is only a preferred embodiment of the present invention, and is not intended to limit the scope of the present invention.

Claims

1. A nucleic acid detecting apparatus, comprising:

the cracking piece is internally provided with a cracking cavity, and is provided with a first through hole communicated with the cracking cavity;

the liquid transferring piece is internally provided with a liquid transferring cavity, the liquid transferring piece is provided with a second through hole and a third through hole which are communicated with the liquid transferring cavity, and the second through hole is communicated with the first through hole;

the amplification piece is internally provided with an amplification cavity, the amplification cavity is provided with a fourth through hole communicated with the amplification cavity, and the fourth through hole is communicated with the third through hole;

the conducting piece is at least partially arranged in the pipetting cavity and can rotate around a first direction so as to simultaneously close the second through hole and the third through hole and open one of the second through hole and the third through hole; the conducting piece can reciprocate along a first direction to input and output liquid of the liquid transferring cavity according to a set amount;

and the limiting part is arranged on the inner wall of the liquid transferring part so as to limit the displacement of the conducting part in the first direction.

2. The nucleic acid detecting device according to claim 1, wherein the conducting member includes a small diameter portion and a first large diameter portion extending in the first direction and connected to each other, the first large diameter portion being located near a bottom end of the liquid transfer chamber with respect to the small diameter portion; one end of the first large diameter part close to the small diameter part is abutted to the inner wall of the liquid transferring piece, one end of the first large diameter part far away from the small diameter part is provided with an opening communicated with the liquid transferring cavity, and the opening can be rotatably communicated with the second through hole or the third through hole; a space is provided between the small diameter portion and the inner wall.

3. The nucleic acid detecting apparatus according to claim 2, wherein the stopper is a protrusion protruding from the inner wall of the pipetting member, the protrusion being located at a distance from the bottom end of the pipetting chamber larger than the length of the first large diameter portion in the first direction, and the protrusion being spaced from or in contact with the small diameter portion.

4. The nucleic acid detecting apparatus according to claim 3, wherein the projection is provided circumferentially around the inner wall of the pipetting member.

5. The nucleic acid detecting device according to claim 2, wherein the opening extends in the first direction, and a length of the opening in the first direction is longer than distances from the second through hole and the third through hole to a bottom end of the pipetting cavity.

6. The nucleic acid detecting apparatus according to claim 5, wherein a distance from the second through hole to a bottom end of the pipetting cavity is the same as a distance from the third through hole to a bottom end of the pipetting cavity.

7. The nucleic acid detecting device according to claim 5, wherein a ratio of a cross-sectional area of the opening to a cross-sectional area of the first large diameter portion is less than 1/10.

8. The nucleic acid detecting apparatus according to claim 2, wherein the conducting member further comprises:

the second large-diameter part extends along the first direction and is connected to one end, far away from the first large-diameter part, of the small-diameter part, and the second large-diameter part protrudes out of the liquid moving cavity.

9. The nucleic acid detecting device according to claim 8, wherein a cross-sectional area of the second large diameter portion is the same as a cross-sectional area of an end of the first large diameter portion near the small diameter portion.

10. The nucleic acid detecting apparatus according to claim 1, wherein the cleavage member and the amplification member are located on both sides of the pipetting member, respectively.