CN114657049B

CN114657049B - Cassette for nucleic acid amplification

Info

Publication number: CN114657049B
Application number: CN202210515294.6A
Authority: CN
Inventors: 蒋太交; 耿鹏; 李胜光; 张辉; 梁松松; 马然
Original assignee: Guangzhou National Laboratory
Current assignee: Guangzhou National Laboratory
Priority date: 2022-05-12
Filing date: 2022-05-12
Publication date: 2023-06-13
Anticipated expiration: 2042-05-12
Also published as: CN114657049A

Abstract

The disclosed embodiments disclose a cartridge for nucleic acid amplification, comprising: the device comprises a cartridge body and a chamber air pressure balancing device, wherein the cartridge body comprises a plurality of first chambers and a cartridge base, the first chambers are arranged on the cartridge base, a first plunger cavity is arranged on the cartridge base, and each of the first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger cylinder and a first plunger, a first plunger cylinder liquid path is arranged on the first plunger cylinder, and the first plunger cylinder can move along the longitudinal axis direction of the first plunger cavity; the chamber pressure balancing device balances the pressure in the plurality of first chambers. According to the cartridge provided by the disclosure, multiple nucleic acid elution can be realized, the loss of magnetic beads in the transfer process is avoided, the phenomenon of liquid mixing between the liquid stored in advance in each chamber can be avoided in the transportation process, the air pressure balance can be realized through the chamber air pressure balance device, and the transfer of liquid is facilitated.

Description

Cassette for nucleic acid amplification

Technical Field

The present disclosure relates to the technical field of biomedical apparatuses, and in particular, to a cartridge for nucleic acid amplification.

Background

The PCR reaction is also called polymerase chain reaction, and is a molecular biological technical means for amplifying specific nucleic acid fragments. PCR technology is widely used in life sciences such as genomic cloning, DNA sequencing, gene expression, medicine, etc. The greatest feature of PCR is that by using a polymerase and primers, nucleic acid amplification can be performed almost in an unlimited amount by using a minute amount of nucleic acid in the instrument. Almost all of the current nucleic acid assays use this method, which is rapid and accurate, and does not last more than two hours from the time the sample is taken to the time the result is obtained. PCR has become the most dominant means of detecting coronaviruses since the new coronaoutbreak. At present, a plurality of cassettes or kits for nucleic acid amplification exist, such as the prior art provides a plunger type nucleic acid amplification cassette, but only single nucleic acid elution can be realized, quantification is difficult, and air is difficult to accurately transfer in the transfer process of liquid.

Disclosure of Invention

In order to solve the problems in the related art, the present disclosure provides a cartridge for nucleic acid amplification.

The cartridge in the present disclosure includes: the device comprises a cartridge body and a chamber air pressure balancing device, wherein the cartridge body comprises a plurality of first chambers and a cartridge base, the first chambers are arranged on the cartridge base, a first plunger cavity is arranged on the cartridge base, and each of the first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger cylinder and a first plunger, a first plunger cylinder liquid path is arranged on the first plunger cylinder, and the first plunger cylinder can move along the longitudinal axis direction of the first plunger cavity; the chamber pressure balancing device balances the pressure in the plurality of first chambers. According to the nucleic acid amplification cartridge provided by the disclosure, multiple nucleic acid elution can be realized, the loss of magnetic beads in the transfer process is avoided, meanwhile, the phenomenon of liquid mixing between the liquid stored in each chamber in advance in the transportation process can be avoided, in addition, the air pressure balance can be realized between each chamber through the chamber air pressure balance device, and the transfer of liquid is facilitated.

As an example, the plurality of first chambers includes a sample loading chamber, a waste chamber, a lysate chamber, at least one wash chamber, an eluent chamber, and a mixing chamber.

As an example, a magnetic element receiving space is provided between the mixing chamber and the first plunger cavity.

As an example, the chamber air pressure balancing device comprises an air passage plate and a puncture plate, wherein an air passage channel and a puncture cavity are arranged on the air passage plate, the air passage channel is communicated with the puncture cavity to form an air passage channel, and a puncture piece on the puncture plate can move up and down along the puncture cavity.

As an example, an isolation seal is provided between the first chamber and the puncture chamber; the piercing member pierces the isolation seal by moving downward through the piercing chamber to communicate the chamber with the gas path channel.

As an example, the isolation seal is provided on the top of the first chamber or on the bottom surface of the gas panel.

Illustratively, the piercing member further includes a seal disposed thereon.

Illustratively, the sealing element comprises a ring-shaped element, wherein grooves matched with the number of the ring-shaped element are formed in the penetrating element, and the ring-shaped element is matched in the grooves.

As an example, the chamber air pressure balancing device comprises an air passage plate, and an air passage is arranged on the top surface of the air passage plate.

As an example, the chamber pressure balancing apparatus further includes a gas switching valve to control transfer of gas from one chamber to another chamber through the gas path channel.

As an example, the gas switching valve includes a plunger disposed below the gas circuit board and a seal ring disposed at a top end of the first chamber, wherein a plunger gas flow path of the plunger is in communication with the chamber when the plunger moves downward to embed the seal ring at the top end of the first chamber.

As an example, the plunger gas flow path includes a through hole provided in a sidewall of the plunger and a blind hole provided in the plunger, wherein the through hole communicates with the blind hole to form a gas flow path.

As an example, the cartridge further comprises a travel control element to control the movement of the plunger.

As an example, the stroke control element includes a cylinder disposed on the cartridge body, and the air circuit board is provided with a through hole to be matched with the cylinder.

As an example, the column includes a first column and a second column with a gap therebetween.

As an example, the cylinder is provided with a first stroke control element and a second stroke control element.

As an example, the rowThe control element comprises a valve arranged on the bottom surface of the gas circuit board "

The convex structure comprises a first flange and a second flange, a gap is reserved between the first flange and the second flange, and a cavity is arranged between at least two adjacent chambers in the plurality of chambers of the first chamber.

As an example, the cavity is provided with at least one set of flange receiving grooves on an inner wall thereof.

As an example, a sealing cover is arranged on the gas circuit board to seal the sample loading chamber.

As an example, an air inlet channel is provided on the cartridge body, and the air inlet channel is communicated with the first plunger cavity.

As an example, the cartridge further comprises a collection device in fluid communication with the first plunger chamber through a collection device fluid path.

As an example, the outer surface of the first plunger barrel is provided with a sealing shell.

As an example, the cartridge body further comprises a plurality of second chambers, a second plunger chamber, wherein each of the second chambers is in fluid communication with the second plunger chamber through a separate second chamber fluid path; a second plunger assembly, wherein the second plunger assembly comprises a second plunger barrel and a second plunger, the second plunger barrel being movable relative to a longitudinal axis of the second plunger barrel; and a second plunger cylinder liquid path is arranged on the wall of the second plunger cylinder.

As an example, the second plunger is provided with a dosing reservoir, and the first plunger chamber and the second plunger chamber are in communication via a transition fluid path provided on the cartridge base.

As an example, the plurality of second chambers includes a secondary reagent chamber and a sealed reagent chamber.

As an example, the sealing agent is stored in advance in the sealing agent chamber, and the sealing agent is at least one of mineral oil, silicone oil, fluoroalkyl oil, vegetable oil, and liquid paraffin.

As an example, a second plunger liquid path is provided on the second plunger, and an overflow flow passage is provided on the second plunger chamber.

As an example, the first plunger and the second plunger may move synchronously.

As an example, the cartridge base may further be provided with an exhaust passage, one end of which communicates with the first plunger chamber and the other end of which communicates with the collecting device.

As an example, the cartridge base is further provided with a push-back liquid path, which communicates with the first plunger chamber and the second plunger chamber, respectively.

As an example, the side wall of the secondary reagent chamber is provided with a secondary reagent sampling tube, and the secondary reagent sampling tube is connected with a sealing plug.

It is to be understood that both the foregoing general description and the following detailed description are exemplary and explanatory only and are not restrictive of the disclosure.

Drawings

Other features, objects and advantages of the present disclosure will become more apparent from the following detailed description of non-limiting embodiments, taken in conjunction with the accompanying drawings. The following is a description of the drawings.

Fig. 1 shows a schematic structural view of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 2 shows an axial view of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 3A shows a schematic structural view of a first plunger barrel of a cartridge for nucleic acid amplification according to the present disclosure.

FIG. 3B is a block diagram showing another embodiment of a first plunger barrel of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 4 shows a schematic structural view of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 5 shows a schematic view of the structural details of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 6 shows a puncture diagram of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

FIG. 7 is a schematic structural view showing still another embodiment of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 8 shows a schematic structural view of a stroke element of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 9 shows a schematic structural view of a cylinder of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

FIG. 10 shows still another schematic structural view of a cylinder of a chamber air pressure balancing device of a cartridge for nucleic acid amplification according to the present disclosure.

FIG. 11 shows still another schematic configuration of the stroke element of the chamber air pressure balancing device of the cartridge for nucleic acid amplification according to the present disclosure.

Fig. 12 shows an assembly schematic view of still another structure of the stroke element of the chamber air pressure balancing device of the cartridge for nucleic acid amplification according to the present disclosure.

Fig. 13 shows a schematic structural view of still another embodiment of a cartridge for nucleic acid amplification according to the present disclosure.

FIG. 14 shows a schematic diagram of an assembled structure of still another embodiment of a cartridge for nucleic acid amplification according to the present disclosure.

Fig. 15 shows a schematic structural view of a second plunger assembly of a cartridge for nucleic acid amplification according to the present disclosure.

FIG. 16 is a schematic view showing a quantitative state structure of still another embodiment of the nucleic acid amplification cartridge of the present disclosure.

It should be understood that the dimensions of the various elements shown in the figures are not drawn to actual scale. Further, the same or similar reference numerals denote the same or similar members.

Detailed Description

Hereinafter, exemplary embodiments of the present disclosure will be described in detail with reference to the accompanying drawings so that those skilled in the art can easily implement them. In addition, for the sake of clarity, portions irrelevant to description of the exemplary embodiments are omitted in the drawings.

In this disclosure, it should be understood that terms such as "comprises" or "comprising," etc., are intended to indicate the presence of features, numbers, steps, acts, components, portions, or combinations thereof disclosed in this specification, and are not intended to exclude the possibility that one or more other features, numbers, steps, acts, components, portions, or combinations thereof are present or added.

The same or similar reference numerals in the drawings of the present disclosure correspond to the same or similar components; in the description of the present disclosure, it should be understood that, if there are terms such as "center", "upper", "lower", "left", "right", "horizontal", "inner", "outer", etc., that indicate orientations or positional relationships based on the orientations or positional relationships shown in the drawings, it is merely for convenience in describing the present disclosure and simplifying the description, rather than indicating or implying that the apparatus or elements being referred to must have a specific orientation, be constructed and operated in a specific orientation, so that the positional relationships described in the drawings are for exemplary illustration only and are not to be construed as limitations of the present disclosure, and that the specific meanings of the above terms may be understood by those of ordinary skill in the art in view of the specific circumstances. Furthermore, the terms "first," "second," "third," and the like, are used merely for distinguishing descriptions of parts and not as indicating or implying a relative importance.

In the description of the present disclosure, it should also be noted that, unless explicitly stated and limited otherwise, the terms "disposed," "mounted," "connected," and "connected" are to be construed broadly, and may be, for example, fixedly connected, detachably connected, or integrally connected; can be mechanically or electrically connected; can be directly connected or indirectly connected through an intermediate medium, and can be communication between two elements. The specific meaning of the terms in this disclosure will be understood by those of ordinary skill in the art in the specific context.

In addition, it should be noted that, without conflict, the embodiments of the present disclosure and features of the embodiments may be combined with each other. The present disclosure will be described in detail below with reference to the accompanying drawings in conjunction with embodiments.

As mentioned above, the nucleic acid amplification apparatus of the prior art has problems that only single nucleic acid elution is achieved, and simultaneously quantification is difficult, and air is difficult to accurately transfer during transfer of liquid, and thus the present disclosure provides a cartridge for nucleic acid amplification to solve the above problems.

According to the present disclosure, there is provided a cartridge for nucleic acid amplification, comprising: the device comprises a cartridge body and a chamber air pressure balancing device, wherein the cartridge body comprises a plurality of first chambers and a cartridge base, the first chambers are arranged on the cartridge base, a first plunger cavity is arranged on the cartridge base, and each of the first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger cylinder and a first plunger, a first plunger cylinder liquid path is arranged on the first plunger cylinder, and the first plunger cylinder can move along the longitudinal axis direction of the first plunger cavity; the chamber pressure balancing device balances the pressure in the plurality of first chambers. According to the nucleic acid amplification cartridge provided by the disclosure, multiple nucleic acid elution can be realized, the loss of magnetic beads in the transfer process is avoided, meanwhile, the phenomenon of liquid mixing between the liquid stored in each chamber in advance in the transportation process can be avoided, in addition, the air pressure balance can be realized between each chamber through the chamber air pressure balance device, and the transfer of liquid is facilitated.

As shown in fig. 1, 2 and 3A, a cartridge 100 for nucleic acid amplification, the cartridge 100 comprises a cartridge body 1 and a chamber air pressure balancing device 2, wherein the cartridge body 1 comprises a plurality of first chambers 10 and a cartridge base 11, the plurality of first chambers 10 are arranged on the cartridge base 11, a first plunger cavity 111 is arranged on the cartridge base 11, and each of the plurality of first chambers 10 is communicated with the first plunger cavity 111 through an independent first chamber liquid path; and a first plunger assembly 3, the first plunger assembly 3 including a first plunger barrel 31 and a first plunger 32, the first plunger barrel 31 being provided with a first plunger barrel fluid path, the first plunger barrel 31 being movable along a longitudinal axis 1111 of the first plunger chamber 111; wherein the chamber pressure balancing means 2 balances the pressure within the plurality of first chambers 10.

As shown in fig. 2, the plurality of first chambers 10 includes a sample loading chamber 101, a waste liquid chamber 102, a lysate chamber 103, at least one wash liquid chamber 104, an eluent chamber 105, and a mixing chamber 106, and as an example, two wash liquid chambers, a first wash liquid chamber 1041 and a second wash liquid chamber 1042, respectively, are shown in fig. 1-2, although those skilled in the art will understand that 1 wash liquid chamber or more wash liquid chambers are possible. Wherein the lysis solution chamber 103, the at least one washing solution chamber 104, and the elution solution chamber 105 store a lysis solution, a washing solution, and an elution solution for nucleic acid extraction, respectively, and the mixing chamber 106 stores a magnetic bead solution in advance. The cartridge body 1 is provided with a plurality of independent liquid paths corresponding to the sample application chamber 101, the waste liquid chamber 102, the lysis liquid chamber 103, the at least one washing liquid chamber 104, the elution liquid chamber 105 and the mixing chamber 106, respectively, so as to communicate each of the aforementioned chambers with the first plunger chamber 111. Specifically, as shown in fig. 2, the liquid path corresponding to the sample application chamber 101 is a sample application chamber liquid path 1011, the liquid path corresponding to the waste liquid chamber 102 is a waste liquid chamber liquid path 1021, the liquid path corresponding to the lysate chamber 103 is a lysate chamber liquid path 1031, the liquid path corresponding to the first wash chamber 1041 is a first wash chamber liquid path 10411, the liquid path corresponding to the second wash chamber 1042 is a second wash chamber liquid path 10421, the liquid path corresponding to the eluent chamber 105 is an eluent chamber liquid path 1051, and the liquid path corresponding to the mixing chamber 106 is a mixing chamber liquid path 1061. In the present disclosure, each liquid path may be formed on the outer surface of the cartridge body 1 by micro-nano processing, and then may be formed by bonding a sealing film on the surface of the cartridge body 1.

As an example, as shown in fig. 2, in the present disclosure, all the ports of the fluid paths of the first chambers on the cartridge body 1 are on a straight line, wherein the straight line is defined along the longitudinal axis direction 1111 of the first plunger chamber 111. In the present disclosure, all of the liquid paths are micron-sized liquid paths, which can be formed by micro-nano processing. As shown in fig. 2-3A, the first plunger barrel 31 is provided with a first plunger barrel liquid path 311, and the first plunger barrel liquid path 311 may be formed by providing a through hole communicating with the first plunger barrel cavity on the side wall of the first plunger barrel. When the cartridge is not in use, the first plunger barrel fluid path 311 is not aligned with any of the fluid paths of the first chambers, and all of the fluid paths of the first chambers are sealed by the first plunger barrel wall, thereby preventing fluid pre-existing within the first chambers from flowing out. When the liquid in the first chamber is required to be transferred, the first plunger barrel 31 can move back and forth along the direction of the longitudinal axis 1111 of the first plunger chamber 111 under the action of external force, so that the first plunger barrel liquid path 311 can be aligned with the port of the liquid path of any one first chamber, after the alignment, the first plunger 32 is pulled back again, the liquid in the aligned first chamber is pumped into the plunger barrel, then the first plunger barrel 31 is moved again, the first plunger barrel liquid path 311 is aligned with the port of the liquid path of the required first chamber, the first plunger 32 is pushed, and the liquid in the first plunger barrel 311 is transferred to the required chamber. Illustratively, the sample solution in the sample application chamber 101 is transferred into the mixing chamber 106 by taking as an illustration that, first, the first plunger barrel 31 is moved along the longitudinal axis of the first plunger cavity 111 by an external force such as a motor, so that the first plunger barrel liquid path 311 is aligned with the liquid outlet of the sample application chamber liquid path 1011, then the first plunger 32 is pulled backward to suck the sample solution in the sample application chamber 101 into the first plunger barrel 31, at this time, the liquid outlet of the liquid path of the remaining first chamber is sealed by the wall of the first plunger barrel 31, the liquid in the chamber cannot flow out, and then the first plunger barrel 31 is moved backward so that the first plunger barrel liquid path 311 is aligned with the liquid outlet of the mixing chamber liquid path 1061, and the sample solution in the first plunger barrel 31 is transferred into the mixing chamber 106 by pushing the first plunger 32. If the repeated suction is needed, the repeated suction can be realized by repeating the steps for a plurality of times.

As shown in fig. 1-2, a magnetic element receiving space 12 may also be provided between the mixing chamber 106 and the first plunger cavity 111 to facilitate magnetic attraction of the magnetic element to the magnetic beads within the mixing chamber 106. By providing a magnetic element receiving space between the mixing chamber 106 and the first plunger cavity 111, space is fully utilized and the structure of the device is reduced.

Fig. 4-6 illustrate a first embodiment of a chamber pressure balancing apparatus.

Referring to fig. 4, the chamber air pressure balancing device 2 includes an air passage plate 21 and a puncture plate 22, wherein the air passage plate 21 is provided with an air passage channel 211 and a plurality of puncture cavities 212, the air passage channel 211 is a micro-sized channel, and as an example, an air passage channel can be formed on the surface of the air passage plate 21 by micro-nano processing, and then the air passage channel can be formed by attaching a film to seal the air passage channel. Each puncture lumen 212 communicates with the gas path channel 211, and in particular, as shown in fig. 5, each puncture lumen 212 communicates with the gas path channel 211 through an intermediate channel 2121. Piercing member 221 on piercing plate 22 is movable up and down along piercing chamber 212 by an external force.

The chamber air pressure balancing device 2 is mounted on the cartridge, specifically, referring to fig. 4, the air circuit board 21 is mounted on the top end of the cartridge body 1, and each puncture cavity 212 corresponds to a chamber other than the sample loading chamber 101, such as the waste liquid chamber 102, the lysis liquid chamber 103, at least one washing liquid chamber 104, the eluent chamber 105 and the mixing chamber 106, in the first chamber. By way of example, isolation seals, such as sealing membranes or the like (not shown), are provided between the waste liquid chamber 102, the lysate chamber 103, the at least one wash liquid chamber 104, the eluent chamber 105, and the mixing chamber 106 and the puncture chamber 212, by means of which the required liquids can be stored in advance in the respective chambers during production without cross-contamination. As an example, the sealing separator may be an aluminum film or a PP film. As an example, the isolation seal may be provided at the top of the first chamber, or the isolation seal may be provided at the bottom surface of the gas circuit board 21.

As mentioned above, piercing member 221 on piercing plate 22 is movable up and down along piercing chamber 212 by an external force. When the puncture member 221 moves downwards along the puncture cavity 212, the puncture member can puncture the sealing spacer when moving to a certain position, and at the moment, a part between the tip of the sealing spacer around the puncture member 221 and the tip of the puncture member 221 have a tiny gap to allow gas to permeate, so that the first chamber is communicated with the gas path channel 211, the gas in each chamber of the first chamber can flow to the user, the gas pressure in each chamber is balanced, and the transfer of liquid is facilitated.

Illustratively, the piercing member is provided with at least one channel to facilitate gas transfer. Specifically, the piercing member 221 has a piercing rod 2211 and a piercing tip 2212 disposed at an end of the piercing rod 2211, and the piercing tip 2212 may be tapered, and at least one groove (not shown) is disposed along a radial direction of the tapered piercing tip 2212 to facilitate transfer of gas from the first chamber into the gas path channel 211 through the groove during piercing.

Illustratively, each piercing member 221 is further provided with a seal 222, where the seal 222 seals the piercing member after the piercing member 221 enters the piercing member 212 to prevent leakage of gas from the piercing site. Specifically, fig. 6 shows the state where the penetration member 221 penetrates the isolation seal assembly. The seal 222 has a diameter that matches the diameter of the first chamber to seal the first chamber. By way of example, as shown in fig. 4-6, the seal 222 may include rings, 2 rings being shown, but those skilled in the art will appreciate that the number of rings may be other numbers, such as 1, 3, or more. The piercing rod 2211 is provided with grooves 22111 matching the number of ring members, each ring member being adapted in the groove 22111, and when the ring member is adapted in the groove 22111, as shown in fig. 6, the diameter thereof is slightly larger than the diameter of the piercing rod so as to seal the first chamber.

Fig. 7-9 show a second embodiment of the chamber pressure balancing device.

As shown in fig. 7-8, the chamber air pressure balancing device 4 includes an air passage plate 41, and an air passage 411 is provided on the top surface of the air passage plate 41. The chamber pressure balancing apparatus also includes a gas switching valve to control the transfer of gas from one chamber to another chamber through gas path 411. Specifically, the gas switching valve includes a plunger 42 and a sealing ring 43, the plunger 42 is disposed on the bottom surface of the gas circuit board 41, the sealing ring 43 is disposed on the top end of the first chamber, and when the plunger 42 moves downward to embed the sealing ring 43 on the top end of the first chamber and moves to a specified position, a plunger gas flow path 421 of the plunger 42 is in communication with the first chamber. The plunger gas flow path 421 is communicated with the gas path channel 411 through the intermediate flow path 412, so that the gas path channel 411 can be communicated with each chamber of the first chamber, gas transfer between each chamber is realized, and finally, the air pressure balance in each chamber is achieved, so that liquid transfer is facilitated. As an example, as shown in fig. 7, the plug gas flow path 421 includes a plug through hole 4211 provided at a side wall of the plug and a blind hole 4212 provided at the plug, wherein the plug through hole 4211 communicates with the blind hole 4212 to form the plug gas flow path 421, wherein an open end of the blind hole 4212 communicates with the intermediate flow path 412.

In this embodiment, unlike the first embodiment of the chamber air pressure balancing device, the air passage 411 on the air passage plate 41 is not connected to each plunger 42, the plunger corresponding to the sample addition chamber 101 is not provided with a plunger air passage 421, the cartridge body 1 is provided with a second air passage 15 to connect the sample addition chamber 101 to the waste liquid chamber 102, accordingly, referring to fig. 7, the plunger corresponding to the sample addition chamber 101 is provided with a plunger air passage 422 of the sample addition chamber, the plunger corresponding to the waste liquid chamber 102 is provided with a plunger air passage 423 of the waste liquid chamber, the

plunger air passages

422 and 423 of the waste liquid chamber can be formed by processing a through hole in the plunger communicating with the plunger cavity, and the plunger air passage 423 of the waste liquid chamber is provided above the plunger through hole 4211, and the plunger air passage 422 and the plunger air passage 423 of the waste liquid chamber can connect the sample addition chamber 101 to the waste liquid chamber 102 through the second air passage 15 to maintain the air pressure balance therebetween. As mentioned above, when the plunger 42 moves down to the seal ring 43 fitted into the top end of the first chamber and moves to the specified position, the plunger gas flow path 421 of the plunger 42 is communicated with the first chamber, and the sample addition chamber plunger gas flow path 422 and the waste liquid chamber plunger gas flow path 423 are respectively communicated with the second gas path 15, whereby the pressure between the sample addition chamber 101 and the waste liquid chamber 102 can be balanced. Compared with the first embodiment of the chamber air pressure balancing device, the first embodiment has simple process and lower cost.

As an example, the cartridge may further comprise a travel control element to control the movement of the plunger, in particular to control the formation of the downward movement of the plunger.

Fig. 8-9 illustrate one embodiment of a travel control element. As shown in fig. 8, the stroke control member includes a cylinder 13 provided on the cartridge body 1, and a through hole 413 is provided on the air passage plate 41 to cooperate with the cylinder 13 to control the stroke of the downward movement of the plunger 42. Specifically, the through hole 413 is sleeved on the column 13, the diameter of the column 13 is slightly larger than the diameter of the through hole 413 or the diameter of the column 13 is matched with the diameter of the through hole 413, but the friction force between the two is larger than the weight of the air circuit board, so that when no downward pressure is applied to the air circuit board 41, the air circuit board 41 is fixed on the column 13, at this time, the plunger through hole 4211 of the plunger 42 is located above the lowest end 431 of the sealing ring 43, the sealing ring 43 seals the plunger through hole 4211, thereby sealing the first chamber, and the gas in the first chamber cannot be transferred. When downward pressure is applied to the air plate 41, the air plate 41 can move downward along the column 13, so that the plunger through hole 4211 of the plunger 42 is located below the lowest end 431 of the sealing ring 43, and at this time, the sample loading chamber plunger gas flow path 422 and the waste liquid chamber plunger gas flow path 423 are also respectively communicated with the second air path 15, so that the conduction between the first chamber and the air path channel 411 and the conduction between the sample loading chamber 101 and the waste liquid chamber 102 are realized, and gas can be transferred from one first chamber to the other chamber, and finally, air pressure balance is realized. It should be noted that, although fig. 8 shows the column 13 disposed between each two first chambers, those skilled in the art will understand that one column may be disposed, for example, in the middle, or 2 or more columns may be uniformly disposed.

As an example, as shown in fig. 8 to 9, in order to facilitate the installation of the air circuit board 41 on the cylinder 13 through the through hole 413, the cylinder 13 may be configured to include a first cylinder 131 and a second cylinder 132, a gap 133 is provided between the first cylinder 131 and the second cylinder 132, and the top ends of the first cylinder 131 and the second cylinder 132 are provided with inward circular-arc-shaped shrinkage smoothing surfaces 134 to facilitate the sleeving of the through hole 413 on the cylinder 13. Specifically, the through holes 413 are aligned with the columns, and since the top ends of the first and

second columns

131 and 132 are provided with the inward circular-arc-shaped shrinkage smooth surfaces 134, the top ends of the first and

second columns

131 and 132 can smoothly enter the through holes 413, and then pressure is applied to the air circuit board 41, and since the gaps 133 are provided between the first and

second columns

131 and 132, the first and

second columns

131 and 132 are close to each other, so that the air circuit board 41 can conveniently move downward along the columns 13.

As an example, as shown in fig. 10, in order to precisely control the downward moving distance of the air circuit board 41, the first stroke control element 135 and the second stroke control element 136 may be further disposed on the cylinder 13 to precisely control the downward moving distance of the air circuit board 41, and as an example, the first stroke control element 135 and the second stroke control element 136 may be ring-shaped members disposed on the cylinder 13; to facilitate insertion of the through-holes into the posts 13, the end face of the first row of control elements 135 is provided with an inwardly rounded, constricted smooth surface 1351. Specifically, referring to fig. 1-2 and 7-10, when the through hole 413 on the air board 41 is sleeved on the first row of control elements 135, the plunger through hole 4211 of the plunger 42 is located above the lowest end 431 of the sealing ring 43, and the sealing ring 43 seals the plunger through hole 4211, so that the first chamber is sealed, and the gas in the first chamber cannot be transferred. When downward pressure is applied to the air plate 41, the air plate 41 can move downward along the column 13 until the through hole 413 is sleeved on the second stroke control element 136, and when the air plate 41 is fixed on the cartridge body, the plunger through hole 4211 of the plunger 42 is located below the lowest end 431 of the sealing ring 43, so that conduction between the first chamber and the air passage 411 is realized, gas can be transferred from one first chamber to the other chamber, and air pressure balance is finally realized.

Fig. 11-15 show yet another embodiment of the travel control element. As shown in FIG. 11, the stroke control member includes a "disposed on the bottom surface of the air circuit board 41"

The "shaped protrusion 414, specifically, the protrusion 414 includes a first flange 4141 and a second flange 4142, a gap is provided between the first flange 4141 and the second flange 4142, and a cavity is provided between at least two adjacent chambers in the plurality of chambers in the first chamber. As shown in fig. 12, a cavity 14 is provided between any two chambers of the plurality of chambers of the first chamber on the cartridge body 1 for receiving the protrusion structure 414 on the air circuit board 41, and when in use, the width of the protrusion structure 414 is slightly larger than the width of the cavity 14 when not deformed, so that the protrusion structure 414 can be buckled in the cavityThe air circuit board 41 can be installed on the cartridge body 1 in the cavity 14, at this time, the plunger through hole 4211 of the plunger 42 is located above the lowest end 431 of the sealing ring 43, and the sealing ring 43 seals the plunger through hole 4211, so that the first cavity is sealed, and the gas in the first cavity cannot be transferred; when downward pressure is applied to the gas circuit board 41, the gas circuit board 41 can move downward along the cavity 14, so that the plunger through hole 4211 of the plunger 42 is located below the lowest end 431 of the sealing ring 43, thereby realizing the conduction between the first chamber and the gas circuit channel 411, and further realizing the transfer of gas from one first chamber to the other, and finally realizing the air pressure balance.

As an example, referring to fig. 12, in order to precisely control the downward moving distance of the air circuit board 41, at least one set of flange receiving grooves is provided on the inner wall of the cavity 14, the flange receiving grooves include a first set of flange receiving grooves 141, and as an example, the first set of flange receiving grooves 141 includes two receiving grooves symmetrically provided at both sides of the cavity 14 for receiving the first flange 4141 and the second flange 4142. When the gas circuit board 41 is used, the width of the protruding structure 414 is slightly larger than the width of the cavity 14 when the protruding structure 414 is not deformed, so that the protruding structure 414 can be buckled in the cavity 14, namely the gas circuit board 41 can be installed on the cartridge body 1, at the moment, the plunger through hole 4211 of the plunger 42 is positioned above the lowest end 431 of the sealing ring 43, the sealing ring 43 seals the plunger through hole 4211, the first cavity is sealed, and gas in the first cavity cannot be transferred; when downward pressure is applied to the air plate 41, the air plate 41 may move downward along the cavity 14, and when the first flange 4141 and the second flange 4142 move to the first group of flange receiving grooves 141, the first flange 4141 and the second flange 4142 may be respectively snapped into the two receiving grooves of the first group of flange receiving grooves 141 due to the outward tension of the first flange 4141 and the second flange 4142, thereby stably mounting the air plate 41 on the cartridge body 1, and at this time, the plunger through hole 4211 of the plunger 42 is located below the lowermost end 431 of the sealing ring 43, thereby achieving the conduction between the first chamber and the air passage 411, so that the transfer of air from one first chamber to the other chamber, and the final air pressure balance can be achieved.

As an example, the first set of flange receiving grooves 141 may be continuously disposed along the inner wall of the cavity 14 so that the first set of flange receiving grooves 141 can smoothly receive the first flange 4141 and the second flange 4142 when the air circuit board is slightly deformed, and the first set of flange receiving grooves 141 and the second set of flange receiving grooves 142 may be continuously disposed along the wall of the cavity 14 to form annular grooves when the cavity 14 is a cylindrical cavity, or may form square grooves when the cavity is square.

As an example, the flange receiving grooves may be two sets, and specifically, referring to fig. 12, the first set of flange receiving grooves 141 and the second set of flange receiving grooves 142, the principle of the second set of flange receiving grooves 142 is the same as that of the first set of flange receiving grooves, and will not be described herein. When in use, the first flange 4141 and the second flange 4142 can be snapped into the first set of flange receiving grooves 141, thereby firmly mounting the air circuit board 41 on the cartridge body 1, and at this time, the plunger through hole 4211 of the plunger 42 is located above the lowest end 431 of the sealing ring 43, the sealing ring 43 seals the plunger through hole 4211, thereby sealing the first chamber, and the gas in the first chamber cannot be transferred; when downward pressure is applied to the air plate 41, since the first flange 4141 and the second flange 4142 may be deformed, and the contact surfaces of the first flange 4141 and the second flange 4142 with the first set of flange receiving grooves 141 have the first flange inclined surface 41411 and the second flange inclined surface 41421, respectively, so that the first flange 4141 and the second flange 4142 may withdraw from the first set of flange receiving grooves 141 to continue to move downward, and when moving to the second set of flange receiving grooves 142, the first flange 4141 and the second flange 4142 may be engaged into the second set of flange receiving grooves 142, thereby stably mounting the air plate 41 on the cartridge body 1, and the plunger through hole 4211 of the plunger 42 is located below the lowermost end 431 of the seal ring 43, thereby achieving the conduction between the first chamber and the air passage 411, thereby enabling the transfer of air from one first chamber to the other, and finally achieving the air pressure balance.

Although shown in FIGS. 10-11 as disposed between any two adjacent plungers 42 "

The "shaped protrusion 414 and any two adjacent first chambers have a cavity 14 therebetween, but those skilled in the art will also appreciate that the protrusion 414 and the corresponding cavity 14 may be one or more.

As an example, a sealing cover is provided on the air circuit board to seal the sample application chamber. As a specific embodiment, as shown in fig. 4, a sealing plug 213 is disposed on the air board 21, a sample loading chamber cavity 214 is correspondingly disposed on the air board 21, when the air board 21 is mounted on the cartridge body 1, the sample loading chamber cavity 214 is in communication with the sample loading chamber 101, and the sample is loaded through the opening of the sample loading chamber cavity 214, and after the sample is loaded, the sealing plug 213 seals the opening of the sample loading chamber cavity 214 to seal the sample loading chamber 101, so as to prevent sample leakage. In particular, the sealing plug 213 is provided with a filter element, which is a breathable waterproof filter element, so that gas can pass through but liquid cannot pass through.

As another specific embodiment, referring to fig. 11, the air circuit board 41 is provided with a sealing cover 415, at this time, the air circuit board 41 is not provided with a sample loading chamber cavity, but directly loads samples from the opening of the sample loading chamber 101, and after loading, the sealing cover 415 is directly sealed with the opening of the sample loading chamber 101. The seal cap 415 also has the same cartridge structure as the seal plug 213.

As an example, with continued reference to fig. 1, an air inlet passage 112 may also be provided on the cartridge base 11, the air inlet passage 112 communicating with an air passage 1121 provided on the cartridge base 11. By providing the air inlet passage 112, air can be introduced into the cartridge device to dry the mixing chamber 106. Specifically, when it is desired to dry the mixing chamber 106 to remove the solvent in the chamber, the first plunger barrel 31 is pulled so that the first plunger barrel 31 moves along the longitudinal axis 1111 of the first plunger cavity 111, the first plunger barrel liquid path 311 is aligned with the gas path 1121, then the first plunger 32 is pulled back to draw clean gas into the first plunger barrel 31, the first plunger barrel 31 is pulled again so that the first plunger barrel liquid path 311 of the first plunger barrel 31 is aligned with the mixing chamber liquid path 1061 corresponding to the mixing chamber 106, the first plunger barrel 31 is pushed again, the gas is pushed into the mixing chamber 106, then the mixing chamber 106 is heated so that the gas is heated to volatilize the residual solvent in the mixing chamber 106, after a period of time, the first plunger 32 is pulled back so that the gas containing the solvent is introduced into the first plunger barrel 31, then the first plunger barrel 31 is pushed so that the first plunger barrel liquid path 311 is aligned with the liquid outlet of the sample loading chamber liquid path (the sample loading chamber is empty), and the first plunger 32 is pushed so that the first plunger barrel 31 contains the solvent 101 into the sample loading chamber 101. As mentioned above, the air circuit board 21 is provided with the sealing plug 213 to seal the sample loading chamber cavity 214, thereby sealing the sample loading chamber 101, or the air circuit board 41 is provided with the sealing cover 415 to seal the sample loading chamber, both the sealing plug 213 and the sealing cover 415 are provided with the air-permeable waterproof filter element, so that the gas with solvent can be discharged from the filter element, and the gas which is not polluted can be filtered and discharged through the filter element. Although the intake passage 112 and the air passage 1121 are shown in fig. 1 as being disposed below the first plunger chamber 111, those skilled in the art will appreciate that the intake passage 112 and the air passage 1121 may be disposed above the first plunger chamber 111. As an example, the inlet passage 112 may also be provided with a filter element to ensure clean gas enters the cartridge.

As an example, referring to fig. 1, the cartridge further includes a collection device 17, and the collection device 17 may be a test tube or a PCR tube, etc., and the collection device 17 communicates with the first plunger chamber 111 through a collection device fluid path 171, so that the eluent in the cartridge may be collected in the collection device. Specifically, when the eluent is collected in the collecting device 17, the first plunger barrel 31 is moved along the longitudinal axis of the first plunger cavity 111 under the action of an external force such as a motor, so that the first plunger barrel liquid path 311 is aligned with the liquid outlet of the mixing chamber liquid path 1061, then the first plunger 32 is pulled backwards to suck the eluent in the mixing chamber 106 into the first plunger barrel 31, at this time, the liquid outlet of the liquid path of the remaining first chamber is sealed by the wall of the first plunger barrel 31, the liquid in the chamber cannot flow out, and then the first plunger barrel 31 is moved backwards to align the first plunger barrel liquid path 311 with the collecting device liquid path 171, so that the eluent in the first plunger barrel 31 can be transferred into the collecting device 17 by pushing the first plunger 32.

As an example, referring to fig. 3B, the outer surface of the first plunger barrel 31 is provided with a seal shell 33, and the seal shell 33 may be made of a soft material that deforms, such as rubber, and by providing the seal shell 33, the bonding effect between the first plunger barrel 31 and the first plunger chamber 111 is better, and leakage of liquid is prevented.

In order to achieve integration of nucleic acid extraction and amplification, referring to fig. 13 to 15, the cartridge body 1 further includes a plurality of second chambers 16; the cartridge base 11 is provided with a second plunger cavity 116, wherein each of the second chambers 16 is in fluid communication with the second plunger cavity 116 via a separate second chamber fluid path;

a second plunger assembly 5, wherein the second plunger assembly 5 comprises a second plunger barrel 51 and a second plunger 52, the second plunger barrel 51 being movable in a direction relative to a longitudinal axis 1161 of the second plunger cavity 116; a second plunger liquid path 512 is provided on the wall of the second plunger 51. In the disclosure, the second plunger liquid path 512 may be formed by providing a through hole in a sidewall of the second plunger, which communicates with the second plunger cavity, and the first chamber 10 is disposed in line with the second chamber 16. By providing the second plunger cavity 116, it is ensured that there is no intersection of the liquid in the second chamber 16 with the liquid in the first chamber 10, avoiding affecting the biochemical reaction. As an example, as shown in fig. 14 to 16, the first plunger assembly 3 and the second plunger assembly 5 are provided on both sides of the cartridge body 1, respectively. In use, after the eluent is transferred to the collection device, reagents such as amplification reagents, sealing reagents, etc. in the second chamber 16 can be transferred into the second plunger barrel 51 through the second chamber fluid path, and then the second plunger barrel 51 is moved such that the second plunger barrel fluid path 512 is aligned with the collection device fluid path 171, and amplification reagents, sealing reagents, etc. are transferred into the collection device 17 for amplification reactions, etc.

Illustratively, the wall of the second plunger 51 is further provided with a dosing reservoir 511, the first plunger chamber 111 and the second plunger chamber 116 being in communication via a transition fluid path 113 provided in the cartridge base 11, the dosing reservoir 511 being adapted for dosing the fluid. Specifically, a groove may be formed on the outer wall of the second plunger barrel 51 by micro-nano machining, wherein the inner wall of the second plunger barrel 51 forms the bottom of the groove, thereby forming the dosing tank 511.

As an example, the second plunger barrel 51 is similar to the first plunger barrel, and the outer surface thereof may be provided with a sealing shell as well, and the manner in which the sealing shell is provided and the function of which is referred to the sealing shell of the first plunger barrel will not be described herein.

It has been mentioned above that liquid may be transferred from one chamber of the first chamber to the other. During nucleic acid extraction, the eluate containing the nucleic acid is eventually within the mixing chamber 106 of the first chamber. In the present embodiment, the first plunger chamber 111 and the second plunger chamber 116 communicate through a transition liquid passage 113. The nucleic acid eluent in the mixing chamber 106 can be introduced into the quantitative tank 511 through the transition liquid path 113. Specifically, the first plunger barrel 31 is moved so that the first plunger barrel liquid path 311 is aligned with the corresponding mixing chamber liquid path 1061 of the mixing chamber 106, then the first plunger 32 is pulled backward to suck the nucleic acid eluent in the mixing chamber 106 into the first plunger barrel 31, at this time, the liquid outlet of the liquid path of the remaining first chamber is sealed by the wall of the first plunger barrel 31, the liquid in the chamber cannot flow out, then the first plunger barrel 31 is moved backward so that the first plunger barrel liquid path 311 is aligned with the liquid inlet of the transition liquid path 113 (i.e., toward the opening of the first plunger cavity 111), the second plunger barrel 51 is moved at the same time, the quantitative pool 511 is aligned with the liquid outlet of the transition liquid path 113 (i.e., toward the opening of the second plunger cavity 116), and then the first plunger 32 is pushed so that the nucleic acid eluent in the first plunger barrel 31 enters into the quantitative pool 511. It should be noted that, according to the inner diameter of the first plunger barrel 31 and the volume of the dosing tank 511, the pushing distance of the first plunger 32 may be accurately determined under the action of an external motor to ensure that the dosing tank is filled with the liquid.

As an example, the plurality of second chambers 16 includes a secondary reagent chamber 161 and a sealed reagent chamber 162, the plurality of second chamber fluid circuits includes a secondary reagent chamber fluid circuit 1611, a sealed reagent chamber fluid circuit 1621, and both the secondary reagent chamber 161 and the sealed reagent chamber 162 are respectively in communication with the second plunger chamber 116 through the secondary reagent chamber fluid circuit 1611 and the sealed reagent chamber fluid circuit 1621, wherein the secondary reagent chamber fluid circuit 1611 and the sealed reagent chamber fluid circuit 1621 are independent fluid circuits. In the present disclosure, the ports of the second chamber on the cartridge body 1 are all in a straight line, wherein the straight line is defined in a direction along the longitudinal axis 1161 of the second plunger chamber 116. The secondary reagent chamber 161 is used for adding secondary reagents such as amplification reagents and the like, and the sealing reagent chamber 162 is pre-stored with sealing reagents, wherein the sealing reagents are at least one of mineral oil, silicone oil, fluorocarbon oil, vegetable oil and liquid paraffin, and can be used for sealing liquid and pushing the liquid in the quantitative tank to drive the liquid to transfer, and the sealing reagents are pre-stored and then sealed by a piston. In particular, it has been mentioned above that the nucleic acid eluate has been transferred into the quantification reservoir 511, and further transfer into the collection device 17 as mentioned above is required, and it will be appreciated by a person skilled in the art that in case a second plunger is provided, the collection device 17 is correspondingly arranged to communicate with the second plunger 116 via the collection device liquid path 171 for facilitating the transfer of liquid into the collection device. When it is desired to transfer the nucleic acid eluent in the quantitative pool 511 into the collection device 17, the second plunger cylinder 51 is moved so that one end of the quantitative pool 511 is aligned with the sealing reagent chamber liquid path 1621 and the other end is aligned with the collection device liquid path 171, and then the plunger of the sealing reagent chamber 162 is controlled to move downward, such as by motor pushing, so that the sealing reagent is transferred into the quantitative pool 511 through the sealing reagent chamber liquid path 1621, thereby pushing the nucleic acid eluent to be transferred into the collection device 17 through the collection device liquid path 171. By this transfer mode, the residue of the nucleic acid eluent can be avoided.

As an example, referring to fig. 13, a vent channel 114 may be further provided on the cartridge base 11, where one end of the vent channel 114 communicates with the first plunger cavity 111 and one end communicates with the collection device 17, so that during transferring the nucleic acid eluent from the dosing tank 511 to the collection device 17, the gas in the collection device 17 may be transferred into the first plunger barrel 31 to facilitate pushing the nucleic acid eluent by the sealing agent into the collection device. Specifically, when transferring the nucleic acid eluting solution from the quantitative reservoir 511 to the collection device 17, the first plunger barrel liquid path 311 of the first plunger barrel 31 is aligned with the air discharge passage 114 by the synchronous operation, and then the first plunger is pulled synchronously, whereby the gas in the collection device 17 can be transferred into the first plunger barrel 31, and the air pressure in the collection device 17 can be reduced to facilitate the elution solution to enter the collection device 17.

As an example, with continued reference to fig. 13-16, the second plunger cavity 116 is provided with an overflow channel 1162, and the formation of the overflow channel 1162 may refer to the formation of the first chamber channel. The end of the dosing tank 511 close to the second plunger liquid path 512 is located at a distance L from the second plunger liquid path 512, and the overflow path 1162 has the same length as L. By providing the overflow channel 1162, it is possible to ensure that the eluent can be filled in the quantitative tank 511 to be accurately quantified, and accurate quantification can be achieved without calculating the distance by which the first plunger is pushed according to the inner diameter of the first plunger cylinder or the like. Specifically, referring to fig. 16, when the eluting solution is transferred from the mixing chamber 106 to the dosing tank 511, the eluting solution of the mixing chamber 106 may be transferred into the first plunger barrel 31 in the above-described manner, and then the first plunger barrel 31 may be moved so that the first plunger barrel liquid path 311 is aligned with the transition liquid path 113 while the second plunger barrel 51 is moved so that the dosing tank 511 is aligned with the transition liquid path 113, and at this time, one end of the overflow channel 1162 is aligned with the dosing tank 511 and the other end is aligned with the second plunger barrel liquid path 512, whereby when the nucleic acid eluting solution is transferred to the dosing tank 511, the first plunger 32 is pushed while the second plunger 52 is pulled at the same rate as the first plunger so that the first plunger and the second plunger move synchronously, so that the nucleic acid eluting solution overflows the dosing tank 511, and the overflowed nucleic acid eluting solution enters into the second plunger barrel 51 through the second plunger barrel liquid path 512, and at this time, the nucleic acid eluting solution completely fills the dosing tank 511, whereby the distance pushed by the first plunger is not required to be precisely controlled. Simultaneously, the first plunger is simultaneously pushed and pulled with the second plunger, in this disclosure, synchronous push and pull movement means that when one plunger is pushed forward at a rate V, the other plunger is pulled back at the same rate V. Through synchronous push-pull movement, on one hand, the liquid to be transferred can be guided; on the other hand, because the air exists in the plunger cavity, the liquid is transferred simply by the driving force or the attractive force of a single plunger, the liquid is difficult to transfer cleanly and can be remained in the runner, and the liquid can be transferred more fully by synchronous push-pull movement without being remained in the pipetting runner. The method for transferring the eluent in the quantifying tank 511 to the collecting device 17 is the same as that described above, and will not be described here.

As an example, referring to fig. 13, the cartridge base 11 is further provided with a push-back liquid path 115, and the push-back liquid path 115 communicates with the first plunger chamber 111 and the second plunger chamber 116, respectively. As mentioned above, in transferring the eluent to the quantitative pool 511, the nucleic acid overflowed from the quantitative pool is sucked into the second plunger barrel 51, the nucleic acid eluent in the second plunger barrel 51 can be pushed back into the first plunger barrel through the push-back liquid path 115, specifically, after the eluent in the quantitative pool 511 is transferred into the collecting device 17, the first plunger barrel liquid path 311, the second plunger barrel liquid path 512 are aligned with the push-back liquid path 115, and then the second plunger 52 is pushed, and simultaneously the first plunger 32 is pulled, so that the eluent in the second plunger barrel 51 can be transferred into the first plunger barrel 31. The eluate in the first plunger barrel may then be transferred to the mixing chamber 106 as needed.

13-16, the secondary reagent chamber 161 is used to add a secondary reagent, such as an amplification reagent, and it is noted that the secondary reagent chamber is sealed with a piston after the addition of the amplification reagent. When the amplification reagent is added, as described above, the eluent in the second plunger barrel 51 has been transferred into the first plunger barrel 31, and at this time the second plunger barrel 51 is empty, the amplification reagent added into the secondary reagent chamber 161 can be transferred into the collection device 17 through the second plunger barrel 51 for the amplification reaction. Specifically, after the eluent in the second plunger barrel 51 is transferred to the first plunger barrel 31, the second plunger barrel 51 is moved so that the second plunger barrel liquid path 512 is aligned with the second reagent chamber liquid path 1611, then the piston sealing the second reagent chamber is pushed down, and the second plunger 52 is pulled at the same time so that the amplification reagent is transferred into the second plunger barrel 51, then the second plunger barrel 51 is moved so that the second plunger barrel liquid path 512 is aligned with the collection device liquid path 171, and the first plunger barrel 31 is moved so that the first plunger barrel liquid path 311 is aligned with the exhaust channel 114, and then the second plunger 52 is pushed, and then the first plunger 32 is pulled at the same time, so that the second amplification reagent is transferred into the nucleic acid collection device 17 with the nucleic acid eluent collected therein for amplification, and in this process, the gas in the collection device 17 can be transferred into the first plunger barrel, thereby reducing the gas pressure in the collection device 17, so that the amplification reagent is convenient to enter the collection device 17. If multiple rounds of amplification are needed, the steps are repeated to finish the multiple rounds of amplification.

As an example, the side wall of the secondary reagent chamber 161 is provided with a secondary reagent sampling tube 1612, the secondary reagent sampling tube 1612 is connected with a secondary reagent sampling tube sealing plug 16121, and when a secondary reagent is added, the secondary reagent can be also added through the secondary reagent sampling tube 1612, and after the addition is completed, the sealing is performed by the secondary reagent sampling tube sealing plug 16121.

It should be noted that, during production, the chamber air pressure balancing device is installed on the cartridge body, but the chambers are not communicated through the air paths, that is, the air pressures in the chambers are respectively kept independent, and the air pressures of the chambers are balanced in the use process.

As mentioned above, the lysis solution chamber 103, the at least one washing solution chamber 104 and the elution solution chamber 105 store a lysis solution, a washing solution and an elution solution for nucleic acid extraction, respectively, while the mixing chamber 106 stores a magnetic bead solution in advance. The principle of use of the present disclosure is as follows:

the method comprises the following steps:

s1, balancing the air pressure of the first chamber: the air pressure in each first cavity is balanced through the cavity air pressure balancing device, and the specific balancing mode is referred to in the mode described above;

s2, activating magnetic beads: the magnetic bead solution in the mixing chamber 106 is pushed into the first plunger barrel 31 according to the liquid transferring mode, and then the first plunger 32 is pushed to enable the magnetic bead solution to return to the mixing chamber 106, and the magnetic bead is activated by repeated pumping;

S3, magnetic bead waste liquid discharge: after the magnetic beads are activated, moving the magnetic elements to the magnetic element accommodating space 12 to fix the magnetic beads in the mixing chamber 106, then transferring all the solution into the first plunger barrel 31, and then transferring the liquid in the first plunger barrel 31 into the waste liquid chamber 102;

s4, adding a sample: adding a sample to the sample adding chamber 101, transferring the sample solution into the first plunger barrel 31, and then transferring the sample solution in the first plunger barrel 31 into the mixing chamber 106;

s5, cleavage reaction: transferring the pyrolysis liquid pre-stored in the pyrolysis liquid chamber 103 into the first plunger barrel 31, and then transferring the pyrolysis liquid in the first plunger barrel 31 into the mixing chamber 106 for pyrolysis reaction; after the pyrolysis liquid is transferred into the mixing chamber 106, repeated suction can be performed by repeatedly pushing and pulling the first plunger 32, so that the pyrolysis reaction is sufficient; after the reaction is completed, moving the magnetic element to the magnetic element accommodating space 12 to fix the magnetic beads in the mixing chamber 106, transferring the waste pyrolysis liquid generated after the reaction is completed into the first plunger barrel 31, and transferring the waste pyrolysis liquid in the first plunger barrel 31 into the pyrolysis liquid chamber 103;

s6, washing: the pre-stored washing liquid in the washing liquid chamber 104 is transferred to the first plunger barrel 31, then the washing liquid in the first plunger barrel 31 is transferred to the mixing chamber 106, and after the washing liquid is transferred to the mixing chamber 106, repeated suction can be performed by repeatedly pushing and pulling the first plunger 32, so that the washing is sufficient. After the washing is sufficient, the magnetic element is moved to the magnetic element accommodating space 12 to fix the magnetic beads in the mixing chamber 106, then the washing liquid waste liquid is transferred to the first plunger barrel 31, and then the washing liquid waste liquid in the first plunger barrel 31 is transferred to the cracking liquid chamber 103;

S7, eluting: transferring the eluent pre-stored in the eluent chamber 105 to the first plunger barrel 31, transferring the eluent in the first plunger barrel 31 to the mixing chamber 106, and repeatedly sucking the eluent by repeatedly pushing and pulling the first plunger 32 after the eluent is transferred to the mixing chamber 106, so that the eluent is fully eluted;

s8, collecting eluent: after the elution is completed, the magnetic element is moved to the magnetic element accommodating space 12 to fix the magnetic beads in the mixing chamber 106, then the eluent is transferred into the first plunger barrel 31, and then the eluent in the first plunger barrel 31 is transferred into a collection device such as a PCR tube.

In the case where the quantitative pool 511 is provided, after the completion of the elution, the magnetic element is moved to the magnetic element accommodating space 12 to fix the magnetic beads in the mixing chamber 106, then the elution is transferred to the first plunger barrel 31, then the elution in the first plunger barrel 31 is transferred to the quantitative pool 511, and then the elution in the quantitative pool 511 is driven into the collection device 17 by the sealing agent. The quantification process is described with reference to the foregoing.

Where an overflow channel 1162 is also provided, this may also include transferring eluent that enters the second plunger barrel 51 via overflow channel 1162 into the mixing chamber 106 via push-back fluid path 115, as described above.

As an example, there may also be a step S0 ventilation between steps S6 and S7: clean gas is introduced into the mixing chamber 106 through the gas inlet channel 112 (see above for the way of introducing the gas into the mixing chamber 106), then the mixing chamber 106 is heated to ensure that the gas is heated to evaporate the residual liquid in the mixing chamber, forming a dry environment, and then the gas is discharged through the sample loading chamber 101 (see above for the way of discharging).

When nucleic acid amplification is desired, the following operations may be performed in conjunction with FIGS. 13-16:

s8, sealing eluent by a sealing reagent: transferring the sealing reagent pre-stored in the sealing reagent chamber 162 into the second plunger barrel 51, and then transferring the sealing reagent in the second plunger barrel 51 into the collecting device 17 to seal the eluent;

s8 amplification reaction: the amplification reagent is added to the secondary reagent chamber 161, and then transferred into the second plunger barrel 51, and then the amplification reagent in the second plunger barrel 51 is transferred to the collection device 17 for the amplification reaction.

In the case where the exhaust passage is provided, in the process of transferring the amplification reagent to the collection device 17, the transfer of the gas of the collection device 17 into the second plunger barrel 51 is also included, and the specific process is referred to above.

The foregoing description is only of the preferred embodiments of the present disclosure and description of the principles of the technology being employed. It will be appreciated by those skilled in the art that the scope of the invention referred to in this disclosure is not limited to the specific combination of features described above, but encompasses other embodiments in which any combination of features described above or their equivalents is contemplated without departing from the inventive concepts described. Such as those described above, are mutually substituted with the technical features having similar functions disclosed in the present disclosure (but not limited thereto).

Claims

1. A nucleic acid amplification cartridge comprising:

the cartridge body and the chamber air pressure balancing device,

the cartridge body comprises a plurality of first chambers and a cartridge base, the first chambers are arranged on the cartridge base, a first plunger cavity is arranged on the cartridge base, and each of the first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger cylinder and a first plunger, a first plunger cylinder liquid path is arranged on the first plunger cylinder, and the first plunger cylinder can move along the longitudinal axis direction of the first plunger cavity;

The chamber air pressure balancing device balances air pressures in the first chambers;

wherein,,

the chamber air pressure balancing device comprises an air circuit board, an air circuit channel is arranged on the top surface of the air circuit board,

the chamber pressure equalization device further includes a gas switching valve to control the transfer of gas from one chamber to another chamber through the gas path passage,

the gas switching valve comprises a plunger and a sealing ring, the plunger is arranged below the gas circuit board, the sealing ring is arranged at the top end of the inner part of the cavity of the first cavity, and when the plunger moves downwards to be embedded into the sealing ring at the top end of the first cavity, a plunger gas flow path of the plunger is communicated with the cavity;

the cartridge further includes a travel control element to control movement of the plunger;

the cartridge body further comprises a plurality of second chambers and a second plunger chamber, wherein each of the second chambers is communicated with the second plunger chamber through an independent second chamber liquid path;

a second plunger assembly, wherein the second plunger assembly comprises a second plunger barrel and a second plunger, the second plunger barrel being movable relative to a longitudinal axis of the second plunger barrel; a second plunger cylinder liquid path is arranged on the wall of the second plunger cylinder;

The second plunger cylinder is provided with a quantitative pool, and the first plunger cavity is communicated with the second plunger cavity through a transition liquid path arranged on the cartridge base;

an overflow runner is arranged on the second plunger cavity;

the stroke control element comprises a valve seat arranged on the bottom surface of the gas circuit board "

The novel flange structure comprises a convex structure in a shape, wherein the convex structure comprises a first flange and a second flange, a gap is reserved between the first flange and the second flange, a cavity is arranged between at least two adjacent cavities in a plurality of cavities of the first cavity, and at least one group of flange receiving grooves are arranged on the inner wall of the cavity.

2. The cartridge for nucleic acid amplification according to claim 1, wherein the plunger gas flow path includes a plunger through hole provided in a side wall of the plunger and a blind hole provided in the plunger, and wherein the plunger through hole communicates with the blind hole to form a gas flow path.

3. The cartridge for nucleic acid amplification according to claim 1, wherein the stroke control member comprises a cylinder provided on the cartridge body, and the gas circuit board is provided with a through hole to be engaged with the cylinder.

4. The nucleic acid amplification cartridge of claim 3, wherein the cartridge comprises a first cartridge and a second cartridge with a gap therebetween.

5. The nucleic acid amplification cartridge of claim 3 or 4, wherein the cartridge is provided with a first stroke control element and a second stroke control element.

6. The nucleic acid amplification cartridge of any one of claims 1-4, wherein the plurality of first chambers comprises a sample loading chamber, a waste chamber, a lysate chamber, at least one wash chamber, an eluate chamber, and a mixing chamber.

7. The nucleic acid amplification cartridge of claim 6, wherein a magnetic element accommodating space is provided between the mixing chamber and the first plunger chamber.

8. The nucleic acid amplification cartridge according to any one of claims 1 to 4 and 7, wherein a sealing cover is provided on the gas circuit board to seal the sample addition chamber.

9. The nucleic acid amplification cartridge according to any one of claims 1 to 4 and 7, wherein an air intake passage is provided in the cartridge body, and the air intake passage communicates with the first plunger chamber.

10. The nucleic acid amplification cartridge of any one of claims 1-4 or 7, further comprising a collection device in fluid communication with the first plunger chamber via a collection device fluid path.

11. The nucleic acid amplification cartridge according to any one of claims 1 to 4 or 7, wherein a seal case is provided on an outer surface of the first plunger cylinder.

12. The nucleic acid amplification cartridge of claim 1, wherein the plurality of second chambers comprises a secondary reagent chamber and a sealing reagent chamber.

13. The nucleic acid amplification cartridge according to claim 12, wherein a sealing agent is stored in advance in the sealing agent chamber, and the sealing agent is at least one of mineral oil, silicone oil, fluorocarbon oil, vegetable oil, and liquid paraffin.

14. The nucleic acid amplification cartridge of claim 1, wherein the first plunger and the second plunger are movable in synchronization.

15. The nucleic acid amplification cartridge according to any one of claims 1 to 12 to 14, wherein a vent passage is further provided in the cartridge base, and one end of the vent passage communicates with the first plunger chamber and the other end communicates with the collection device.

16. The cartridge for nucleic acid amplification according to claim 15, wherein the cartridge base is further provided with a back-pushing liquid path which communicates with the first plunger chamber and the second plunger chamber, respectively.

17. The cartridge for nucleic acid amplification according to claim 14, wherein the cartridge base is further provided with a back-pushing liquid path which communicates with the first plunger chamber and the second plunger chamber, respectively.

18. The cartridge for nucleic acid amplification according to any one of claims 12 to 13, wherein a secondary reagent sampling tube is provided on a side wall of the secondary reagent chamber, and a sealing plug is connected to the secondary reagent sampling tube.