CN114657049A

CN114657049A - Card box for nucleic acid amplification

Info

Publication number: CN114657049A
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: 2022-06-24
Anticipated expiration: 2042-05-12
Also published as: CN114657049B

Abstract

The disclosed embodiments disclose a cartridge for nucleic acid amplification, including: the card box comprises a card box body and a chamber air pressure balancing device, wherein the card box body comprises a plurality of first chambers and a card box base, the plurality of first chambers are arranged on the card box base, a first plunger cavity is arranged on the card box base, and each of the plurality of first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger barrel and a first plunger, a first plunger barrel liquid path is arranged on the first plunger barrel, and the first plunger barrel can move along the longitudinal axis direction of the first plunger cavity; the chamber air pressure balancing device balances the air pressure in the plurality of first chambers. According to this box that openly provides, can realize many times nucleic acid elution, avoid the loss of magnetic bead in the transfer process, can avoid taking place the liquid mixing phenomenon between the liquid of prestoring in each cavity in the transportation, can realize atmospheric pressure through cavity atmospheric pressure balancing unit and balance, make things convenient for the transfer of liquid.

Description

Card box for nucleic acid amplification

Technical Field

The disclosure relates to the technical field of biomedical instruments, in particular to a card box for nucleic acid amplification.

Background

The PCR reaction is also called polymerase chain reaction, and is a molecular biological technique for amplifying and amplifying specific nucleic acid fragments. PCR technology is widely used in the field of life sciences, such as genomic cloning, DNA sequencing, gene expression, medicine, etc. The greatest feature of PCR is that nucleic acid amplification can be carried out almost limitlessly using a polymerase and primers and a trace amount of nucleic acid in an instrument. Most of the current methods for nucleic acid detection are rapid and accurate, and the time from taking a sample to obtaining a result is not more than two hours. Since the new outbreak, PCR has become the most prominent means for detecting coronaviruses. At present, a plurality of cartridges or kits for nucleic acid amplification are available, for example, a plunger type nucleic acid amplification cartridge is provided in the prior art, but the cartridge can only realize single nucleic acid elution and is difficult to quantify, and the liquid has the problems that air is difficult to accurately transfer in the transfer process, and the like.

Disclosure of Invention

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

The cartridge in the present disclosure comprises: the cassette comprises a cassette body and a chamber air pressure balancing device, wherein the cassette body comprises a plurality of first chambers and a cassette base, the plurality of first chambers are arranged on the cassette base, a first plunger cavity is arranged on the cassette base, and each of the plurality of first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger barrel and a first plunger, a first plunger barrel liquid path is arranged on the first plunger barrel, and the first plunger barrel can move along the longitudinal axis direction of the first plunger cavity; the chamber air pressure balancing device balances air pressure in the plurality of first chambers. According to this nucleic acid is card box for amplification that this disclosure provides, can realize many times nucleic acid elution, avoid the loss of magnetic bead in the transfer process, can realize avoiding taking place the cluster liquid phenomenon between the liquid of prestoring in each cavity simultaneously in the transportation, can realize atmospheric pressure balance through cavity atmospheric pressure balancing unit between each cavity in addition, make things convenient for the transfer of liquid.

Illustratively, the plurality of first chambers includes a sample addition chamber, a waste chamber, a lysis chamber, at least one wash chamber, an elution chamber, and a mixing chamber.

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

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

As an example, an isolation seal is arranged between the first chamber and the puncture chamber; the puncture piece punctures the isolation sealing piece by moving downwards through the puncture cavity so as to conduct the cavity and the air channel.

Illustratively, the isolation seal is disposed on a top of the first chamber or a bottom surface of the gas panel.

Illustratively, the piercing member is further provided with a sealing member.

Illustratively, the sealing element comprises a ring-shaped element, the puncturing element is provided with grooves with the number matched with that of the ring-shaped element, and the ring-shaped element is matched in the grooves.

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

Illustratively, the chamber air pressure balancing device further comprises a gas switching valve to control the transfer of gas from one chamber to another chamber through the air passage.

Illustratively, 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 first chamber, and when the plunger moves downwards to be embedded into the sealing ring at the top end of the first chamber, a plunger gas flow path of the plunger is communicated with the chamber.

Illustratively, the plunger gas flow path comprises a through hole arranged on the side wall of the plunger and a blind hole arranged on the plunger, wherein the through hole is communicated with the blind hole to form a gas flow path.

Illustratively, the cartridge further comprises a stroke control element to control movement of the plunger.

Illustratively, the stroke control element comprises a cylinder arranged on the cartridge body, and the gas circuit board is provided with a through hole to be matched with the cylinder.

Illustratively, the cylinder includes a first cylinder and a second cylinder with a gap therebetween.

Illustratively, the cylinder has a first stroke control element and a second stroke control element disposed thereon.

Illustratively, the stroke control member includes a member disposed on a bottom surface of the gas circuit board "

The 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 reserved between at least two adjacent chambers in the plurality of chambers of the first chamber.

Illustratively, at least one set of flange-receiving slots is provided on an inner wall of the cavity.

Illustratively, the gas circuit board is provided with a sealing cover to seal the sample application chamber.

Illustratively, an air inlet channel is arranged on the cartridge body and is communicated with the first plunger cavity.

Illustratively, the cartridge further comprises a collection device in fluid communication with the first plunger chamber via a collection device fluid path.

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

Illustratively, the cartridge body further comprises a plurality of second chambers, a second plunger cavity, wherein each of the second chambers is in communication with the second plunger cavity 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 direction of the second plunger cavity; and a second plunger cylinder liquid path is arranged on the cylinder wall of the second plunger.

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

Illustratively, the plurality of second chambers includes a secondary reagent chamber and a sealed reagent chamber.

Illustratively, the sealing agent chamber is pre-stored with a sealing agent, and the sealing agent is at least one of mineral oil, silicone oil, fluorocarbon oil, vegetable oil and liquid paraffin.

As an example, a second plunger cylinder liquid path is arranged on the second plunger cylinder, and an overflow flow passage is arranged on the second plunger cavity.

Illustratively, the first plunger and the second plunger may move synchronously.

As an example, the cartridge base can be further provided with an exhaust channel, one end of the exhaust channel is communicated with the first plunger cavity, and the other end of the exhaust channel is communicated with the collecting device.

Illustratively, the cartridge base is further provided with a push-back fluid circuit, which is in communication with the first and second plunger chambers, respectively.

Illustratively, the side wall of the secondary reagent chamber is provided with a secondary reagent sample adding pipe, and the secondary reagent sample adding pipe 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 when taken in conjunction with the accompanying drawings. The following is a description of the drawings.

Fig. 1 shows a schematic configuration diagram 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 the cartridge for nucleic acid amplification according to the present disclosure.

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

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

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

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

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

FIG. 8 is a schematic view showing the structure of a stroke member of the chamber air pressure equalizing device of the cartridge for nucleic acid amplification according to the present disclosure.

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

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

FIG. 11 is a schematic view showing still another structure of a stroke member of the chamber air pressure equalizing device of the cartridge for nucleic acid amplification according to the present disclosure.

FIG. 12 is a schematic view showing the assembly of still another structure of the stroke member of the chamber air pressure equalizing device of the cartridge for nucleic acid amplification according to the present disclosure.

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

FIG. 14 is a schematic view showing an assembly structure of still another embodiment of the cartridge for nucleic acid amplification according to the present disclosure.

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

Fig. 16 is a schematic diagram showing a quantitative state structure of a nucleic acid amplification cartridge according to still another embodiment of the present disclosure.

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

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. Also, for the sake of clarity, parts not relevant to the description of the exemplary embodiments are omitted in the drawings.

In the present disclosure, it is to be understood that terms such as "including" or "having," etc., are intended to indicate the presence of the disclosed features, numbers, steps, behaviors, components, parts, or combinations thereof, and are not intended to preclude the possibility that one or more other features, numbers, steps, behaviors, components, parts, or combinations thereof may be present or added.

The same or similar reference numbers 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 "center", "upper", "lower", "left", "right", "horizontal", "inner", "outer", etc., indicating orientations or positional relationships based on those shown in the drawings, it is merely for convenience of description and simplicity of description, but does not indicate or imply that the device or element referred to must have a particular orientation, be constructed and operated in a particular orientation, and therefore the positional relationships depicted in the drawings are for illustrative purposes only and are not to be construed as limitations of the present disclosure, as the specific meaning of the terms described above will 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 to distinguish one element from another, and are not to be construed as indicating or implying relative importance.

Throughout the description of the present disclosure, it should also be noted that, unless expressly stated or limited otherwise, the terms "disposed," "mounted," "connected," and "coupled" are to be construed broadly, e.g., as meaning either a fixed connection, a removable connection, or an integral connection; can be mechanically or electrically connected; either directly or indirectly through intervening media, or may be interconnected between two elements. The specific meaning of the above terms in the present disclosure can be understood in specific instances by those of ordinary skill in the art.

It should be further noted that the embodiments and features of the embodiments in the present disclosure may be combined with each other without conflict. 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 device of the prior art has many problems that only a single elution of nucleic acid can be achieved, and that it is difficult to quantify the nucleic acid, and that it is difficult to accurately transfer a liquid by air during the transfer, and the like, 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 cassette comprises a cassette body and a chamber air pressure balancing device, wherein the cassette body comprises a plurality of first chambers and a cassette base, the plurality of first chambers are arranged on the cassette base, a first plunger cavity is arranged on the cassette base, and each of the plurality of first chambers is communicated with the first plunger cavity through an independent first chamber liquid path; the first plunger assembly comprises a first plunger barrel and a first plunger, a first plunger barrel liquid path is arranged on the first plunger barrel, and the first plunger barrel can move along the longitudinal axis direction of the first plunger cavity; the chamber air pressure balancing device balances air pressure in the plurality of first chambers. According to this nucleic acid is card box for amplification that this disclosure provides, can realize many times nucleic acid elution, avoid the loss of magnetic bead in the transfer process, can realize avoiding taking place the cluster liquid phenomenon between the liquid of prestoring in each cavity simultaneously in the transportation, can realize atmospheric pressure balance through cavity atmospheric pressure balancing unit between each cavity in addition, make things convenient for the transfer of liquid.

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

As shown in fig. 2, the plurality of first chambers 10 includes a sample addition chamber 101, a waste solution chamber 102, a lysis solution chamber 103, at least one wash chamber 104, an elution solution chamber 105, and a mixing chamber 106, and illustratively, two wash chambers are shown in fig. 1-2, namely, a first wash chamber 1041 and a second wash chamber 1042, although those skilled in the art will also understand that there may be 1 wash chamber or more wash chambers. Wherein the lysis solution chamber 103, the at least one wash solution chamber 104, and the elution solution chamber 105 respectively store lysis solution, wash solution, and elution solution for nucleic acid extraction, and the mixing chamber 106 previously stores magnetic bead solution. The cartridge body 1 is provided with a plurality of independent liquid paths respectively corresponding to the sample addition 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 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 addition chamber 101 is a sample addition 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 lysis liquid chamber 103 is a lysis liquid chamber liquid path 1031, the liquid path corresponding to the first washing liquid chamber 1041 is a first washing liquid chamber liquid path 10411, the liquid path corresponding to the second washing liquid chamber 1042 is a second washing liquid chamber liquid path 10421, the liquid path corresponding to the elution liquid chamber 105 is an elution liquid 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.

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

As shown in fig. 1-2, a magnetic element accommodating space 12 may be further disposed between the mixing chamber 106 and the first plunger cavity 111, so that the magnetic element magnetically attracts magnetic beads in the mixing chamber 106. By providing a magnetic element accommodating space between the mixing chamber 106 and the first plunger cavity 111, the space is fully utilized, reducing the structure of the device.

Fig. 4-6 illustrate a first embodiment of a chamber air pressure equalization device.

Referring to fig. 4, the chamber air pressure balancing device 2 includes an air channel plate 21 and a puncture plate 22, the air channel plate 21 is provided with an air channel 211 and a plurality of puncture cavities 212, the air channel 211 is a micron-sized channel, as an example, an air channel groove may be formed on the surface of the air channel plate 21 by micro-nano processing, and then the air channel may be formed by attaching a film to the air channel groove. Each puncture lumen 212 communicates with the air passage channel 211, and in particular, as shown in fig. 5, each puncture lumen 212 communicates with the air passage channel 211 through an intermediate channel 2121. The puncture piece 221 of the puncture plate 22 can move up and down along the puncture cavity 212 by being driven by external force.

The chamber air pressure balancing means 2 is mounted on the cartridge, and specifically, in conjunction with fig. 4, the air plate 21 is mounted on the top end of the cartridge body 1, and each puncture chamber 212 corresponds to a chamber other than the sample addition chamber 101 among the first chambers, such as the waste chamber 102, the lysis chamber 103, the at least one wash chamber 104, the elution chamber 105, and the mixing chamber 106. As an example, the waste liquid chamber 102, the lysis liquid chamber 103, the at least one wash liquid chamber 104, the elution liquid chamber 105, and the mixing chamber 106 and the puncture chamber 212 are provided with a separation seal member such as a sealing film or the like (not shown in the drawings), by which a desired liquid can be stored in each chamber in advance at the time of 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 disposed at the top of the first chamber, or the isolation seal may be disposed at the bottom surface of the gas circuit board 21.

As mentioned above, the piercing member 221 of the piercing plate 22 can move up and down along the piercing cavity 212 under the driving of external force. When the puncture piece 221 moves downwards along the puncture cavity 212, the puncture piece can puncture the sealing partition when moving to a certain position, at the moment, a part of the sealing partition between the tips of the puncture piece 221 and the tips of the puncture piece 221 are provided with fine gaps through which gas can penetrate, so that the first cavity is communicated with the gas path channel 211, the gas in each cavity of the first cavity can flow from a user to the user, the gas pressure in each cavity is balanced, and the liquid is convenient to transfer.

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

By way of example, each piercing member 221 is further provided with a seal 222, and the seal 222 can seal the piercing cavity after the piercing member 221 enters the piercing cavity 212, thereby preventing gas from leaking out of the piercing site. Specifically, fig. 6 shows the state in which the piercing member 221 pierces the barrier seal assembly. The diameter of the seal 222 matches the diameter of the first chamber and seals the first chamber. As an example, as shown in fig. 4-6, the seal 222 may include rings, 2 of which are 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 a number of grooves 22111 matching the number of rings, each ring can fit into a groove 22111, and when the rings fit into the grooves 22111, as shown in fig. 6, the diameter of the rings is slightly larger than the diameter of the piercing rod to seal the first chamber.

Fig. 7-9 illustrate a second embodiment of the chamber air pressure equalization means.

As shown in fig. 7 to 8, the chamber air pressure balancing means 4 includes an air passage plate 41, and an air passage channel 411 is provided on a top surface of the air passage plate 41. The chamber gas pressure balancing means also includes a gas switching valve to control the transfer of gas from one chamber to another through the gas passage 411. Specifically, the gas switching valve includes a plunger 42 and a gasket 43, the plunger 42 is disposed on the bottom surface of the gas passage plate 41, the gasket 43 is disposed on the top end of the first chamber, and when the plunger 42 moves downward to be inserted into the gasket 43 on the top end of the first chamber and moves to a predetermined position, the plunger gas passage 421 of the plunger 42 is communicated with the first chamber. The plunger gas flow path 421 is communicated with the gas path 411 through the intermediate flow path 412, so that the gas path 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. Illustratively, as shown in fig. 7, the plunger gas flow path 421 includes a plunger through hole 4211 provided in a sidewall of the plunger and a blind hole 4212 provided on the plunger, wherein the plunger through hole 4211 communicates with the blind hole 4212 to form the plunger gas flow path 421, wherein an open end of the blind hole 4212 communicates with the intermediate flow path 412.

It should be noted that, unlike the first embodiment of the chamber air pressure balancing device, in this embodiment, the air channel 411 of the air channel plate 41 is not communicated with each plunger 42, the plunger corresponding to the sample addition chamber 101 is not provided with the plunger gas flow path 421, but the cartridge body 1 is provided with the second air channel 15 to communicate the sample addition chamber 101 with the waste liquid chamber 102, accordingly, referring to fig. 7, the plunger of the sample addition chamber corresponding to the sample addition chamber 101 is provided with the sample addition chamber plunger gas flow path 422, the plunger of the waste liquid chamber corresponding to the waste liquid chamber 102 is provided with the waste liquid chamber plunger gas flow path 423, the sample addition chamber plunger gas flow path 422 and the waste liquid chamber plunger gas flow path 423 may be formed by processing a through hole communicating with the plunger chamber on the plunger, and the waste liquid chamber plunger gas flow path 423 is provided above the plunger through hole 4211, the sample addition chamber plunger gas flow path 422 and the waste liquid chamber plunger gas flow path 423 may be formed by connecting the sample addition chamber plunger gas flow path 15 with the plunger chamber plunger gas path The chamber 101 communicates with the waste chamber 102 to maintain a pressure balance therebetween. As mentioned above, when the plunger 42 moves downward and is inserted into the gasket 43 at the top end of the first chamber and moves to a predetermined position, the plunger gas channel 421 of the plunger 42 is communicated with the first chamber, and at this time, the plunger gas channel 422 of the sample addition chamber and the plunger gas channel 423 of the waste liquid chamber are respectively communicated with the second gas path 15, so that the pressure between the sample addition chamber 101 and the waste liquid chamber 102 can be balanced. Compared with the first implementation mode of the chamber air pressure balancing device, the implementation mode is simple in process and lower in cost.

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

Fig. 8-9 illustrate one embodiment of a travel control element. As shown in fig. 8, the stroke control element includes a cylinder 13 provided on the cartridge body 1, and the air passage plate 41 is provided with a through hole 413 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 body 13, the diameter of the column body 13 is slightly larger than that of the through hole 413 or the diameter of the column body 13 and that of the through hole 413 are adapted, but a friction force between the two is larger than the weight of the air channel plate, so that when downward pressure is not applied to the air channel plate 41, the air channel plate 41 is fixed on the column body 13, 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, and thus the first chamber is sealed, and gas in the first chamber cannot be transferred. When downward pressure is applied to the gas channel plate 41, the gas channel 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, 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 gas path 15, so that the conduction between the first chamber and the gas path channel 411 and the conduction between the sample loading chamber 101 and the waste liquid chamber 102 are realized, the gas can be transferred from one first chamber to the other chamber, and finally, the gas pressure balance is realized. It should be noted that although fig. 8 shows that the columns 13 are disposed between every two first chambers, it is understood by those skilled in the art that one column may be disposed, for example, at the middle position, or 2 or more columns may be uniformly disposed.

As an example, as shown in fig. 8-9, in order to facilitate the gas channel plate 41 to be installed 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 an inward arc-shaped contracting and smoothing surface 134 to facilitate the through hole 413 to be sleeved on the cylinder 13. Specifically, the through hole 413 is aligned with the cylinders, since the top ends of the first cylinder 131 and the second cylinder 132 are provided with the inward circular arc-shaped contraction smooth surfaces 134, the top ends of the first cylinder 131 and the second cylinder 132 can smoothly enter the through hole 413 and then apply pressure to the gas path plate 41, and since the gap 133 is provided between the first cylinder 131 and the second cylinder 132, the first cylinder 131 and the second cylinder 132 are close to each other, which facilitates the gas path plate 41 to move downward along the cylinder 13.

For example, as shown in fig. 10, in order to precisely control the downward movement distance of the gas circuit board 41, a first stroke control element 135 and a second stroke control element 136 may be further disposed on the column 13 to precisely control the downward movement distance of the gas 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 column 13; to facilitate insertion of the through-hole into the cylinder 13, the end face of the first stroke control element 135 is provided with an inwardly radiused converging smooth surface 1351. Specifically, with reference to fig. 1-2 and 7-10, when the through hole 413 of the air channel plate 41 is sleeved on the first stroke control element 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 channel plate 41, the air channel plate 41 can move downwards along the cylinder 13 until the through hole 413 is sleeved on the second stroke control element 136, at this time, when the air channel 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 the first chamber is communicated with the air channel 411, gas can be transferred from one first chamber to another chamber, and finally, the air pressure balance is realized.

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

The "shaped protrusion 414, specifically, the protrusion 414 includes a first flange 4141 and a second flange 4142, there is a gap between the first flange 4141 and the second flange 4142, and a cavity is disposed between at least two adjacent chambers of the plurality of chambers of the first chamber. As shown in fig. 12, a cavity 14 is provided between any two of the first chamber and the second chamber on the cartridge body 1 for receiving the protruding structure 414 on the air channel plate 41, when in use, the protruding structure 414 has a slightly wider width than the cavity 14 when not deformed, so that the protruding structure 414 can be fastened in the cavity 14, that is, the air channel plate 41 can be mounted on the cartridge body 1, 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 as to seal the first chamber, and the air in the first chamber cannot be transferred; when downward pressure is applied to the air passage plate 41, the air passage plate 41 can move downwards along the cavity 14, so that the plunger through hole 4211 of the plunger 42 is positioned below the lowest end 431 of the sealing ring 43, so that the first chamber is communicated with the air passage channel 411, gas can be transferred from one first chamber to the other chamber, and finally, the air pressure balance is realized.

For example, referring to fig. 12, in order to precisely control the distance that the gas circuit board 41 moves downward, at least one set of flange receiving grooves including a first set of flange receiving grooves 141 is provided on the inner wall of the cavity 14, and the first set of flange receiving grooves 141 includes, for example, two receiving grooves symmetrically provided on both sides of the cavity 14 for receiving the first flange 4141 and the second flange 4142. When the cartridge 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 fastened in the cavity 14, that is, the gas circuit board 41 can be installed on the cartridge body 1, 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, so that the first chamber is sealed, and gas in the first chamber cannot be transferred; when downward pressure is applied to the gas plate 41, the gas plate 41 can move downward along the cavity 14, and when the first and second flanges 4141 and 4142 move to the first set of flange receiving grooves 141, the first and second flanges 4141 and 4142 can be respectively snapped into the two receiving grooves of the first set of flange receiving grooves 141 due to the outward tension of the first and second flanges 4141 and 4142, thereby stably mounting the gas plate 41 on the cartridge body 1, with the plunger through hole 4211 of the plunger 42 located below the lowest end 431 of the sealing ring 43, thereby achieving communication between the first chamber and the gas passage 411, thereby achieving gas transfer from one first chamber to another, and finally achieving gas pressure balance.

For 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 even when the gas 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, or square grooves when the cavity 14 is a cylindrical cavity.

As an example, the flange receiving slots may be two sets, specifically, referring to fig. 12, a first set of flange receiving slots 141 and a second set of flange receiving slots 142, and the principle of the second set of flange receiving slots 142 is the same as that of the first set of flange receiving slots, which is not described herein again. In use, the first and second flanges 4141, 4142 may snap into the first set of flange receiving grooves 141, thereby securely mounting the gas plate 41 to the cartridge body 1 with the plunger through hole 4211 of the plunger 42 above the lowermost end 431 of the sealing ring 43, the sealing ring 43 sealing the plunger through hole 4211, thereby sealing the first chamber, where gas cannot be transferred; when downward pressure is applied to the gas panel 41, since the first flange 4141 and the second flange 4142 can be deformed, and the contact surfaces of the first and second flanges 4141 and 4142 with the first set of flange receiving grooves 141 have first and second flange inclined surfaces 41411 and 41421, whereby the first and second flanges 4141 and 4142 may be withdrawn from the first set of flange receiving grooves 141 to continue moving downward, when moved into the second set of flange receiving slots 142, the first and second flanges 4141, 4142 may again snap into the second set of flange receiving slots 142, thereby, the air channel plate 41 is stably installed 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 packing 43, therefore, the first chamber is communicated with the air channel 411, so that air can be transferred from one first chamber to the other chamber, and air pressure balance is finally realized.

Although it is shown in fig. 10-11 that any two adjacent plungers 42 are disposed therebetween "

The "shaped protrusion structure 414 and any two adjacent first cavities have a cavity 14 therebetween, but those skilled in the art can also understand that there may be one or more protrusion structures 414 and corresponding cavities 14.

Illustratively, a sealing cover is arranged on the gas path plate to seal the sample adding chamber. As a specific embodiment, as shown in fig. 4, a sealing plug 213 is disposed on the gas path plate 21, a sample loading chamber 214 is correspondingly disposed on the gas path plate 21, when the gas path plate 21 is mounted on the cartridge body 1, the sample loading chamber 214 is conducted with the sample loading chamber 101, loading is performed through an opening of the sample loading chamber 214, and after loading is completed, the sealing plug 213 seals the opening of the sample loading chamber 214 to seal the sample loading chamber 101, thereby preventing sample leakage. In particular, the sealing plug 213 is provided with a filter element, which is permeable to air and waterproof and can ensure that gas but not liquid can pass through.

As another specific embodiment, referring to fig. 11, a sealing lid 415 is provided on the gas channel plate 41, in this case, the sample is directly loaded from the opening of the sample loading chamber 101 without providing a sample loading chamber cavity on the gas channel plate 41, and after the sample loading is completed, the opening of the sample loading chamber 101 is directly sealed by the sealing lid 415. The seal 415 also has the same cartridge arrangement as the seal plug 213.

As an example, with continued reference to fig. 1, an air inlet channel 112 may also be provided on the cartridge base 11, and the air inlet channel 112 is communicated with an air channel 1121 provided on the cartridge base 11. By providing the inlet channel 112, gas can be introduced into the cartridge device to dry the mixing chamber 106. Specifically, when the mixing chamber 106 needs to be dried to remove the solvent in the chamber, the first plunger cylinder 31 is pulled to move the first plunger cylinder 31 along the longitudinal axis 1111 of the first plunger cavity 111, the first plunger cylinder liquid path 311 is aligned and communicated with the air path channel 1121, the first plunger 32 is pulled backwards to suck clean air into the first plunger cylinder 31, the first plunger cylinder 31 is pulled again to pull the first plunger cylinder liquid path 311 of the first plunger cylinder 31 to be aligned with the mixing chamber liquid path 1061 corresponding to the mixing chamber 106, the first plunger cylinder 31 is pushed again to push air into the mixing chamber 106, the mixing chamber 106 is heated to volatilize the solvent remaining in the mixing chamber 106, after a certain period of time, the first plunger 32 is pulled backwards to allow the air containing the solvent to enter the first plunger cylinder 31, and the first plunger cylinder 31 is pushed to align the first plunger cylinder liquid path 311 with the liquid outlet of the sample adding chamber 1011 (at this time, the sample adding chamber is an empty chamber) The first plunger 32 is pushed, so that the gas containing the solvent in the first plunger cylinder 31 enters the sample addition chamber 101. As mentioned above, the gas channel plate 21 is provided with the sealing plug 213 to seal the sample adding chamber 214 and thus the sample adding chamber 101, or the gas channel plate 41 is provided with the sealing cover 415 to seal the sample adding chamber, and both the sealing plug 213 and the sealing cover 415 are provided with the gas permeable and waterproof filter element, so that the gas with the solvent can be discharged from the filter element, and the gas which is not polluted can be discharged through the filter element. Although fig. 1 shows the air inlet passage 112 and the air passage 1121 disposed below the first plunger cavity 111, those skilled in the art will appreciate that the air inlet passage 112 and the air passage 1121 may also be disposed above the first plunger cavity 111. Illustratively, the inlet passage 112 may also be provided with a filter element to ensure that clean gas enters the cartridge.

Illustratively, referring to fig. 1, the cartridge further comprises a collection device 17, the collection device 17 may be a test tube or a PCR tube, etc., and the collection device 17 is in communication with the first plunger cavity 111 through a collection device fluid path 171, so that the eluate in the cartridge can be collected in the collection device. Specifically, when the eluent is finally collected in the collecting device 17 in the mixing chamber 106, the first plunger cylinder 31 is moved along the longitudinal axis along the first plunger cavity 111 under the action of an external force, such as a motor, so that the first plunger cylinder liquid path 311 is aligned with the liquid outlet of the mixing chamber liquid path 1061, then the first plunger 32 is pulled backward to suck the eluent in the mixing chamber 106 into the first plunger cylinder 31, at this time, the liquid outlets of the remaining first chamber liquid paths are sealed by the cylinder wall of the first plunger cylinder 31, the liquid in the chambers cannot flow out, and then the first plunger cylinder 31 is moved backward so that the first plunger cylinder liquid path 311 is aligned with the collecting device liquid path 171, so as to push the first plunger 32, thereby transferring the eluent in the first plunger cylinder 31 into the collecting device 17.

As an example, referring to fig. 3B, the outer surface of the first plunger cylinder 31 is provided with a sealing shell 33, the sealing shell 33 may be made of a soft material that is deformed, such as rubber, and the sealing shell 33 may be provided to make the fitting effect between the first plunger cylinder 31 and the first plunger cavity 111 better and prevent liquid leakage.

In order to realize 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 second plunger chambers 116, wherein each of the second chambers 16 is in fluid communication with the second plunger chambers 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 cylinder fluid path 512 is provided in the wall of the second plunger cylinder 51. In the disclosure, the second plunger cylinder fluid path 512 may be formed by providing a through hole in the sidewall of the second plunger cylinder, which is in communication with the second plunger cylinder chamber, and the first chamber 10 and the second chamber 16 are linearly arranged. By providing the second plunger chamber 116, it is ensured that the liquid in the second chamber 16 does not intersect with the liquid in the first chamber 10, thereby avoiding the influence on the biochemical reaction. Illustratively, as shown in fig. 14 to 16, the first plunger assembly 3 and the second plunger assembly 5 are respectively provided on both sides of the cartridge body 1. In use, after the elution solution is transferred to the collection device, reagents such as amplification reagents, sealing reagents and the like in the second chamber 16 can be transferred to the second plunger cylinder 51 through the second chamber liquid path, and then the second plunger cylinder 51 is moved, so that the second plunger cylinder liquid path 512 is aligned with the collection device liquid path 171, and the amplification reagents, the sealing reagents and the like are transferred to the collection device 17 for amplification reaction and the like.

Illustratively, the wall of the second plunger cylinder 51 is further provided with a quantitative pool 511, the first plunger cavity 111 and the second plunger cavity 116 are communicated through a transition liquid path 113 arranged on the cartridge base 11, and the quantitative pool 511 is used for quantitatively transferring liquid. Specifically, a groove may be processed on the outer wall of the second plunger cylinder 51 by micro-nano processing, wherein the inner wall of the second plunger cylinder 51 forms the bottom of the groove, thereby forming the quantification pool 511.

For example, the second plunger barrel 51 is similar to the first plunger barrel, and the outer surface thereof may also be provided with a sealing shell, and the arrangement and function of the sealing shell refer to the sealing shell of the first plunger barrel, which is not described herein again.

As already mentioned, the liquid may be transferred from one chamber of the first chamber to the other chamber. During the nucleic acid extraction process, the nucleic acid-containing eluent is finally 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 with each other through a transition liquid passage 113. The nucleic acid eluent in the mixing chamber 106 can be introduced into the quantitative cell 511 via the transition liquid path 113. Specifically, the first plunger cylinder 31 is moved to align the first plunger cylinder liquid path 311 with the mixing chamber liquid path 1061 corresponding to the mixing chamber 106, then the first plunger 32 is pulled backwards to suck the nucleic acid eluent in the mixing chamber 106 into the first plunger cylinder 31, while the liquid outlets of the liquid paths of the remaining first chambers are sealed by the cylinder wall of the first plunger cylinder 31, so that the liquid in the chambers cannot flow out, and then the first plunger cylinder 31 is moved backwards to align the first plunger cylinder liquid path 311 with the liquid inlet of the transition liquid path 113 (i.e., the opening toward the first plunger cavity 111), while the second plunger cylinder 51 is moved to align the quantitative pool 511 with the liquid outlet of the transition liquid path 113 (i.e., the opening toward the second plunger cavity 116), and then the first plunger 32 is pushed to make the nucleic acid eluent in the first plunger cylinder 31 enter the quantitative pool 511. It should be noted that, according to the inner diameter of the first plunger cylinder 31 and the volume of the quantitative pool 511, the distance that the first plunger 32 pushes can be precisely determined under the action of the external motor to ensure that the quantitative pool is filled with the liquid.

Illustratively, the plurality of second chambers 16 includes a secondary reagent chamber 161 and a sealed reagent chamber 162, the plurality of second chamber fluid paths includes a secondary reagent chamber fluid path 1611 and a sealed reagent chamber fluid path 1621, both the secondary reagent chamber 161 and the sealed reagent chamber 162 are respectively communicated with the second plunger cavity 116 through the secondary reagent chamber fluid path 1611 and the sealed reagent chamber fluid path 1621, and the secondary reagent chamber fluid path 1611 and the sealed reagent chamber fluid path 1621 are independent fluid paths. In the present disclosure, the ports of the fluid path of the second chamber on the cartridge body 1 are all in a line, wherein the line is defined in a direction along the longitudinal axis 1161 of the second plunger cavity 116. The secondary reagent chamber 161 is used for adding a secondary reagent such as an amplification reagent, and the sealing reagent chamber 162 is pre-stored with a sealing reagent, wherein the sealing reagent is 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 pool to drive the liquid to transfer, and the sealing reagent is pre-stored and then sealed by a piston. In particular, where it has been mentioned above that the nucleic acid eluent has been transferred to the quantification pool 511 and needs to be further transferred to the aforementioned collection device 17, it will be appreciated by those skilled in the art that where a second plunger barrel is provided, the collection device 17 is correspondingly arranged to communicate with the second plunger cavity 116 via the collection device fluid path 171 to facilitate the transfer of liquid into the collection device. When the nucleic acid eluent in the quantitative pool 511 needs to be transferred into the collection device 17, the second plunger cylinder 51 is moved first 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 piston of the sealing reagent chamber 162 is controlled to move downwards, such as being pushed by a motor, 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, the remaining of the nucleic acid eluent can be avoided.

As an example, referring to fig. 13, an air exhaust channel 114 may be further disposed on the cartridge base 11, one end of the air exhaust channel 114 is communicated with the first plunger cavity 111, and the other end is communicated with the collection device 17, so that during the process of transferring the nucleic acid eluent from the quantitative pool 511 to the collection device 17, the gas in the collection device 17 can be transferred into the first plunger cylinder 31 to facilitate the sealing reagent to push the nucleic acid eluent into the collection device. Specifically, when the nucleic acid eluent is transferred from the quantitative pool 511 to the collection device 17, the first plunger barrel liquid path 311 of the first plunger barrel 31 is aligned with the exhaust channel 114 by synchronous operation, and then the first plunger is pulled synchronously, so that the gas in the collection device 17 can be transferred into the first plunger barrel 31, and the gas pressure in the collection device 17 is reduced, so that the eluent can conveniently 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 passage 1162, and the formation of the overflow passage 1162 may be referred to the formation of the first chamber passage. A distance L exists between the end part of the quantitative pool 511 close to the second plunger cylinder liquid path 512 and the second plunger cylinder liquid path 512, and the length of the overflow flow passage 1162 is the same as L. By providing the overflow flow channel 1162, the eluent can be ensured to be filled in the quantification pool 511, thereby accurate quantification can be achieved without calculating the distance pushed by the first plunger according to the inner diameter of the first plunger cylinder and the like. Specifically, referring to fig. 16, when the eluent is transferred from the mixing chamber 106 to the quantitative pool 511, the eluent in the mixing chamber 106 may be transferred into the first plunger barrel 31 according to the method mentioned above, and then the first plunger barrel 31 is moved such that the first plunger barrel liquid path 311 is aligned with the transition liquid path 113, and the second plunger barrel 51 is moved such that the quantitative pool 511 is aligned with the transition liquid path 113, while one end of the overflow flow passage 1162 is aligned with the quantitative pool 511, and the other end is aligned with the second plunger barrel liquid path 512, so that when the nucleic acid eluent is transferred into the quantitative pool 511, the first plunger 32 is pushed, and the second plunger 52 is pulled at the same speed as the first plunger, so that the first plunger and the second plunger are moved synchronously, so that the nucleic acid eluent overflows the quantitative pool 511, and the overflowing nucleic acid eluent enters the second plunger barrel 51 through the second plunger barrel liquid path 512, while the nucleic acid eluent completely fills the quantitative pool 511, whereby the distance of the first plunger push need not be precisely controlled. At the same time, a simultaneous push-pull movement of the first plunger with the second plunger, which in this disclosure means that one plunger pushes forward at a rate V while the other plunger pulls backward at the same rate V. By 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 chamber, the liquid is transferred only by the driving force or the attraction force of a single plunger, the liquid is difficult to transfer completely and can remain in the flow channel, and the liquid can be transferred more fully and cannot remain in the liquid transfer flow channel by synchronous push-pull movement. The method for transferring the eluate in the quantification pool 511 to the collection device 17 is the same as that described above, and will not be described herein.

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

As mentioned above, with reference to FIGS. 13-16, the secondary reagent chamber 161 is used for adding secondary reagents such as amplification reagents, and it is noted that the secondary reagent chamber is sealed with a piston after the amplification reagents are added. When the amplification reagent is added, the eluent in the second plunger cylinder 51 is transferred to the first plunger cylinder 31 as described above, and the second plunger cylinder 51 is empty, so that the amplification reagent added to the secondary reagent chamber 161 can be transferred to the collection device 17 through the second plunger cylinder 51 to perform the amplification reaction. Specifically, after the eluent in the second plunger cylinder 51 is transferred to the first plunger cylinder 31, the second plunger cylinder 51 is moved so that the second plunger cylinder fluid path 512 is aligned with the secondary reagent chamber fluid path 1611, then the piston sealing the secondary reagent chamber is pushed down, and at the same time the second plunger 52 is pulled so that the amplification reagent is transferred to the second plunger cylinder 51, then the second plunger cylinder 51 is moved so that the second plunger cylinder fluid path 512 is aligned with the collection device fluid path 171, and at the same time the first plunger cylinder 31 is moved so that the first plunger cylinder fluid path 311 is aligned with the exhaust passage 114, and then the second plunger 52 is pushed, and at the same time the first plunger 32 is pulled synchronously, so that the secondary amplification reagent is transferred to the nucleic acid collection device 17 in which the nucleic acid eluent is collected for amplification, and during which the gas in the collection device 17 is transferred to the first plunger cylinder, thereby reducing the gas pressure in the collection device 17, to facilitate the entry of amplification reagents into the collection means 17. If multiple rounds of amplification are needed, the steps are repeated to complete the multiple rounds of amplification.

Illustratively, a secondary reagent adding tube 1612 is provided on a side wall of the secondary reagent chamber 161, and a secondary reagent adding tube sealing plug 16121 is connected to the secondary reagent adding tube 1612, and when the secondary reagent is added, the secondary reagent can be added through the secondary reagent adding tube 1612, and after the addition is completed, the secondary reagent adding tube sealing plug 16121 is used for sealing.

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 with each other through the air passage, that is, the air pressure in each chamber is kept independent, and the air pressure in each chamber is balanced during use.

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

the method comprises the following steps of:

s1 equalizing the air pressure of the first chamber: balancing the air pressure in each first chamber by the chamber air pressure balancing device, wherein the balancing mode is the mode described in the foregoing;

s2 activated magnetic beads: the magnetic bead solution in the mixing chamber 106 is put into the first plunger cylinder 31 in the liquid transfer manner, and then the first plunger 32 is pushed to return the magnetic bead solution to the mixing chamber 106, and magnetic bead activation is performed by repeated pumping;

s3 discharge of waste magnetic bead liquid: after the magnetic beads are activated, the magnetic element is moved to the magnetic element accommodating space 12 to fix the magnetic beads in the mixing chamber 106, then the solution is completely transferred into the first plunger cylinder 31, and then the liquid in the first plunger cylinder 31 is transferred into the waste liquid chamber 102;

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

s5 cleavage reaction: transferring the lysate prestored in the lysate chamber 103 into the first plunger cylinder 31, and then transferring the lysate in the first plunger cylinder 31 into the mixing chamber 106 for a lysis reaction; after the lysate is transferred into the mixing chamber 106, repeated suction can be performed by repeatedly pushing and pulling the first plunger 32, so that the lysis reaction is sufficient; after the reaction 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 waste lysate generated after the reaction is completely transferred into the first plunger cylinder 31, and then the waste lysate in the first plunger cylinder 31 is transferred into the lysate chamber 103;

s6 washing: the washing liquid pre-stored 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 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 washing liquid waste liquid is transferred into the first plunger cylinder 31, and then the washing liquid waste liquid in the first plunger cylinder 31 is transferred into the lysis liquid chamber 103;

s7 elution: the elution liquid prestored in the elution liquid chamber 105 is transferred to the first plunger barrel 31, then the elution liquid in the first plunger barrel 31 is transferred to the mixing chamber 106, and after the elution liquid is transferred to the mixing chamber 106, the first plunger 32 can be repeatedly pushed and pulled to repeatedly suck so that the elution is sufficient;

s8 collecting the eluate: after 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, and then the eluent is transferred into the first plunger cylinder 31, and then the eluent in the first plunger cylinder 31 is transferred into a collection device such as a PCR tube.

In the case of the quantitative cell 511, after the elution liquid 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 elution liquid is transferred into the first plunger cylinder 31, then the elution liquid in the first plunger cylinder 31 is transferred into the quantitative cell 511, and then the elution liquid in the quantitative cell 511 is driven into the collecting device 17 by using the sealing reagent. The quantification process is described above.

In the case that the overflow flow passage 1162 is further provided, the method further includes transferring the eluent entering the second plunger cylinder 51 through the overflow flow passage 1162 into the mixing chamber 106 through the liquid pushing-back circuit 115, and the specific process is referred to above.

As an example, there may 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 gas into the mixing chamber 106), the mixing chamber 106 is heated to ensure that the gas is heated to evaporate the residual liquid in the mixing chamber, so as to form a dry environment, and then the gas is exhausted through the sample loading chamber 101 (see above for the way of exhausting).

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

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

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

In the case of providing the exhaust channel, the process of transferring the amplification reagents to the collecting device 17 further includes transferring the gas of the collecting device 17 to the second plunger cylinder 51, and the specific process refers to the foregoing.

The foregoing description is only exemplary of the preferred embodiments of the disclosure and is illustrative of the principles of the technology employed. It will be appreciated by those skilled in the art that the scope of the invention in the present disclosure is not limited to the specific combination of the above-mentioned features, but also encompasses other embodiments in which any combination of the above-mentioned features or their equivalents is possible without departing from the inventive concept. For example, the above features and the technical features disclosed in the present disclosure (but not limited to) having similar functions are replaced with each other to form the technical solution.

Claims

1. A cartridge for nucleic acid amplification, comprising:

a card box body and a chamber air pressure balancing device,

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

the chamber air pressure balancing device balances air pressure in the plurality of first chambers.

2. The cartridge for nucleic acid amplification of claim 1, wherein the plurality of first chambers include a sample addition chamber, a waste liquid chamber, a lysis liquid chamber, at least one wash liquid chamber, an elution liquid chamber, and a mixing chamber.

3. The cartridge for nucleic acid amplification according to claim 2, wherein a magnetic element-housing space is provided between the mixing chamber and the first plunger chamber.

4. The cartridge for nucleic acid amplification according to claim 1, wherein the chamber air pressure balancing means comprises an air channel plate and a puncturing plate, the air channel plate is provided with an air channel and a puncturing cavity, the air channel is communicated with the puncturing cavity to form an air channel, and a puncturing member on the puncturing plate is movable up and down along the puncturing cavity.

5. The cartridge for nucleic acid amplification according to claim 4, wherein an isolation seal is provided between the first chamber and the puncture chamber; the puncture piece punctures the isolation sealing piece by moving downwards through the puncture cavity so as to conduct the cavity and the air channel.

6. The cartridge for nucleic acid amplification according to claim 5, wherein the isolation seal is provided on the top of the first chamber or the bottom surface of the gas channel plate.

7. The cartridge for nucleic acid amplification according to any one of claims 4 to 6, wherein the puncture member is further provided with a sealing member.

8. The cartridge for nucleic acid amplification according to claim 7, wherein the sealing member includes a ring-like member, and the puncture member is provided with grooves corresponding in number to the ring-like member, and the ring-like member is fitted in the grooves.

9. The cartridge for nucleic acid amplification according to claim 1, wherein the chamber air pressure balancing means comprises an air channel plate having an air channel provided on a top surface thereof.

10. The cartridge for nucleic acid amplification according to claim 9, wherein the chamber gas pressure balancing means further comprises a gas switching valve to control transfer of gas from one chamber to another chamber through the gas passage channel.

11. The cartridge for nucleic acid amplification according to claim 10, wherein the gas switching valve comprises a plunger disposed below the gas channel plate and a packing disposed at a top end of the first chamber, wherein a plunger gas flow path of the plunger is communicated with the chamber when the plunger moves downward to be fitted into the packing at the top end of the first chamber.

12. The cartridge for nucleic acid amplification according to claim 11, wherein the plunger gas flow path comprises a plunger through hole provided in a side wall of the plunger and a blind hole provided in the plunger, wherein the plunger through hole communicates with the blind hole to form a gas flow path.

13. The cartridge for nucleic acid amplification according to claim 11 or 12, wherein the cartridge further comprises a stroke control element to control movement of the plunger.

14. The cartridge for nucleic acid amplification according to claim 13, wherein the stroke control member comprises a cylinder provided on the cartridge body, and the gas passage plate is provided with a through hole to fit the cylinder.

15. The cartridge for nucleic acid amplification according to claim 14, wherein the column includes a first column and a second column, and a gap is provided between the first column and the second column.

16. The cartridge for nucleic acid amplification according to claim 14 or 15, wherein a first stroke control element and a second stroke control element are provided on the cartridge.

17. The cartridge for nucleic acid amplification according to claim 13, wherein the stroke control member comprises a plate provided on a bottom surface of the gas passage plate "

18. The cartridge for nucleic acid amplification according to claim 17, wherein at least one set of flange receiving grooves is provided on an inner wall of the cavity.

19. The cartridge for nucleic acid amplification according to any one of claims 4 to 6, 8 to 12, 14 to 15, and 17 to 18, wherein a sealing cover is provided on the gas flow plate to seal the sample addition chamber.

20. The cartridge for nucleic acid amplification according to any one of claims 4 to 6, 8 to 12, 14 to 15, and 17 to 18, wherein an air inlet passage is provided in the cartridge body, and the air inlet passage communicates with the first plunger chamber.

21. The cartridge for nucleic acid amplification according to any one of claims 4 to 6, 8 to 12, 14 to 15, and 17 to 18, further comprising a collection device in fluid communication with the first plunger chamber through a collection device fluid path.

22. The cartridge for nucleic acid amplification according to any one of claims 4 to 6, 8 to 12, 14 to 15, and 17 to 18, wherein an outer surface of the first plunger barrel is provided with a sealing shell.

23. The cartridge for nucleic acid amplification according to any one of claims 4 to 6, 8 to 12, 14 to 15, and 17 to 18, wherein the cartridge body further comprises a plurality of second chambers, a second plunger chamber, wherein each of the second chambers communicates with the second plunger chamber through a separate 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 the longitudinal axis of the second plunger chamber; and a second plunger cylinder liquid path is arranged on the cylinder wall of the second plunger.

24. The nucleic acid amplification cartridge according to claim 23, wherein a quantification chamber is provided in the second plunger barrel, and the first plunger chamber and the second plunger chamber are communicated with each other through a transition liquid path provided in a cartridge base.

25. The cartridge for nucleic acid amplification according to claim 24, wherein the plurality of second chambers include a secondary reagent chamber and a sealing reagent chamber.

26. The nucleic acid amplification cartridge of claim 25, wherein a sealing reagent is stored in the sealing reagent chamber in advance, and the sealing reagent is at least one of mineral oil, silicone oil, fluorocarbon oil, vegetable oil, and liquid paraffin.

27. The cartridge for nucleic acid amplification according to claim 24, wherein a second plunger barrel liquid path is provided in the second plunger barrel, and an overflow flow path is provided in the second plunger chamber.

28. The cartridge for nucleic acid amplification according to claim 27, wherein the first plunger and the second plunger are movable in synchronization.

29. The cartridge for nucleic acid amplification of any one of claims 24 to 28, wherein an exhaust channel is further provided on the cartridge base, the exhaust channel having one end communicating with the first plunger chamber and the other end communicating with the collection device.

30. The cartridge for nucleic acid amplification according to claim 27, wherein the cartridge base is further provided with a push-back liquid path, and the push-back liquid path communicates with the first plunger chamber and the second plunger chamber, respectively.

31. The cartridge for nucleic acid amplification according to claim 28, wherein the cartridge base is further provided with a push-back liquid passage, and the push-back liquid passage communicates with the first plunger chamber and the second plunger chamber, respectively.

32. The cartridge for nucleic acid amplification according to any one of claims 25 or 26, wherein a side wall of the secondary reagent chamber is provided with a secondary reagent addition tube to which a sealing plug is connected.