CN115386486A

CN115386486A - Card box and detection device

Info

Publication number: CN115386486A
Application number: CN202110563790.4A
Authority: CN
Inventors: 梅哲; 贾础豪; 张彤
Original assignee: Guangzhou Wondfo Biotech Co Ltd
Current assignee: Guangzhou Wondfo Biotech Co Ltd
Priority date: 2021-05-21
Filing date: 2021-05-21
Publication date: 2022-11-25

Abstract

The invention relates to the technical field of biomedical treatment and discloses a card box and a detection device, wherein the card box comprises a card box body, a reagent pack group and a sealing cover; the main flow channel, the valve group, the reaction cavity and the airflow channel are arranged on the card box body; the reagent pack group is attached to the surface of the card box body and comprises at least four independently sealed reagent pack units; the air flow channels at least comprise a first air flow channel, a second air flow channel, a third air flow channel and a fourth air flow channel which are respectively corresponding to the four reagent pack units, and the test solution in the reagent pack units flows into the corresponding air flow channels after being broken and flows into one or more of the reaction cavities along the main flow channel under the control of the valve group and the air pump. The invention can realize the automatic treatment of nucleic acid extraction, amplification and detection, only needs a few manual operation steps, and has the advantages of simple operation, safety, convenience, strong sealing property and more accurate detection result.

Description

Card box and detection device

Technical Field

The invention relates to the technical field of biomedical treatment, in particular to a card box and a detection device.

Background

Nucleic acid detection plays a very important role in many fields of biochemical analysis, and has been widely applied in the field of biomedical science.

In the prior art, nucleic acid extraction is generally carried out by using a centrifugal column method or a magnetic bead method, and four steps of lysis, combination, washing, elution and the like are generally required. After nucleic acid extraction is carried out on the sample, subsequent steps of nucleic acid molecular hybridization, polymerase Chain Reaction (PCR), biochip and the like are carried out continuously to complete nucleic acid detection.

Due to the more steps of nucleic acid detection, the whole 'from sample to result' full-automatic detection device is very difficult to realize. In addition, as for the transfer of the effective components in each step, a manual transfer mode is adopted in the prior art, so that the operation is complex, time and labor are wasted, the materials are difficult to transfer fully and efficiently, the result is unstable due to manual operation, and the detection difficulty is high.

In addition, the mainstream technology of molecular detection is the fluorescence quantitative PCR technology, and because the PCR technology has the characteristic of exponential amplification template, the existing open consumables easily cause PCR aerosol pollution in the whole operation process and influence the purity of an extract, thereby limiting the further clinical application of the fluorescence quantitative PCR technology.

Thus, improvements in the prior art are needed.

Disclosure of Invention

The purpose of the invention is: the invention provides a card box and a detection device, which are used for solving the technical problems that in the prior art, the full-automatic processing of nucleic acid detection is difficult to realize, and the detection result is unstable, the detection accuracy is low and the detection efficiency is low due to manual operation and open consumable material use.

In order to achieve the above object, the present invention provides a cartridge comprising a cartridge body, a reagent pack group, and a cover;

the card box body is provided with a main flow channel, a valve group, a reaction cavity and an airflow channel; the valve group comprises a plurality of valves arranged at the preset position of the main flow channel, so that one ends of the reaction chamber and the air flow channel are selectively communicated with the main flow channel, and the other end of the air flow channel is provided with an air vent for connecting an air pump;

the reaction cavity comprises a sample cavity, a magnetic bead storage cavity, a magnetic bead capturing cavity, a waste liquid cavity, a mixing cavity and a PCR cavity; the sample cavity is respectively communicated with the magnetic bead storage cavity and the first end of the magnetic bead capturing cavity; the first end of the magnetic bead capturing cavity is also communicated with the mixing cavity, and the opposite second end of the magnetic bead capturing cavity is communicated with the waste liquid cavity; the mixing cavity is communicated with the PCR cavity; the sample cavity is provided with a sample adding port, and the cover is used for sealing the sample adding port;

the reagent pack group is attached to the surface of the card box body and comprises at least four independently sealed reagent pack units;

the air flow channels at least comprise a first air flow channel, a second air flow channel, a third air flow channel and a fourth air flow channel which respectively correspond to the four reagent pack units; after the reagent pack unit is broken, the built-in test solution correspondingly flows into the first air flow channel, the second air flow channel, the third air flow channel and the fourth air flow channel, and flows into one or more of the reaction chambers along the main flow channel under the control of the valve group and the air pump.

In some embodiments of the present application, four of the reagent pack units are a lysis solution pack, a binding solution pack, a first washing solution pack and an elution solution pack corresponding to the first gas flow channel, the second gas flow channel, the third gas flow channel and the fourth gas flow channel, respectively;

the first gas flow channel is communicated with the magnetic bead storage cavity, the second gas flow channel is communicated with the sample cavity, the third gas flow channel is communicated with the sample cavity or the first end of the magnetic bead capturing cavity, and the fourth gas flow channel is communicated with the second end of the magnetic bead capturing cavity.

the first gas flow channel is communicated with the sample cavity, the second gas flow channel is communicated with the magnetic bead storage cavity, the third gas flow channel is communicated with the first end of the sample cavity or the magnetic bead capturing cavity, and the fourth gas flow channel is communicated with the second end of the magnetic bead capturing cavity.

In some embodiments, the reagent pack further comprises a second wash solution pack, and the air channel further comprises a fifth air channel corresponding to the second wash solution pack, wherein the fifth air channel is communicated with the first end of the sample chamber or the magnetic bead capture chamber.

In some embodiments of the present application, a sixth air passage, a seventh air passage and an eighth air passage are further disposed on the cartridge body; one end of the sixth air passage, one end of the seventh air passage and one end of the eighth air passage are respectively provided with the air vent and the air pump, the other end of the sixth air passage is communicated with the top of the sample cavity, the other end of the seventh air passage is communicated with the blending cavity, and the other end of the eighth air passage is communicated with the waste liquid cavity.

In some embodiments of the present application, the sample chamber includes a sample addition chamber and a sample processing chamber, and the sample addition chamber and the sample processing chamber are communicated through a siphon bend; one end of the siphon bent pipe is communicated with the bottom of the sample adding cavity, and the other end of the siphon bent pipe is communicated with the top of the sample processing cavity; the sample adding port is arranged at the top of the sample adding cavity.

In some embodiments of the present application, an internal reference freeze-dried bead chamber is further disposed between the siphon elbow and the sample addition chamber; one end of the internal reference freeze-dried bead cavity is communicated with the bottom of the sample adding cavity, and the other end of the internal reference freeze-dried bead cavity is communicated with the siphon bent pipe.

In some embodiments of the present application, a blocking lumen is further included; the blocking cavity is connected to the main runner between the blending cavity and the PCR cavity, so that the blending cavity is blocked from being communicated with the PCR cavity when a preset condition is met.

In some embodiments of the present application, the blocking chamber is a wax valve chamber, inside which solid wax is pre-disposed.

In some embodiments of the present application, the cartridge body comprises a cartridge plate, a front membrane, and a rear membrane; the front surface of the cassette board is provided with a cavity or a through hole, the front film is attached to the front surface of the cassette board, and the rear film is attached to the back surface of the cassette board to seal the cavity or the through hole, so that the reaction cavity is formed.

In some embodiments of the present application, the main flow channel is composed of at least one positive and negative trough structure; the front groove structure and the back groove structure comprise a front groove, a back groove and a communication hole; the front groove is arranged on the front surface of the cassette plate, the back groove is arranged on the back surface of the cassette plate, and the communication hole communicates the front groove with the back groove; the front and rear films seal the front and rear grooves and the communication hole to form the main flow passage.

In some embodiments of the present application, the PCR chamber comprises a central chamber, an amplification chamber, and a zone flow channel;

the central cavity is communicated with the flow passage; the amplification cavity is provided with at least one, and the amplification cavity is communicated with the central cavity through the region flow channel.

In some embodiments of the present application, the PCR chamber further comprises a central flow channel groove, a gas containing region, a waterproof gas permeable membrane, and a region gas channel;

the central runner groove is arranged on one surface of the central cavity, the runner is arranged on the other surface opposite to the central cavity, the runner is communicated with one end of the central runner groove, and the other end of the central runner groove is communicated with the zone runner; the waterproof breathable film is attached to the central cavity to seal the central runner groove; one end of the air passage is arranged at the edge of the sealing area of the waterproof breathable film, and the other end of the air passage is communicated with the air containing area.

In some embodiments of the present application, two of the region channels are provided for one of the amplification chambers, and the two region channels are an entry region channel and an exit region channel; one end of the entering area flow channel is communicated with the central flow channel groove, and the other end of the entering area flow channel is communicated with one end of the amplification cavity; one end of the outlet flow channel is communicated with the other end of the amplification cavity, and the port of the other end of the outlet flow channel is positioned in the central cavity and sealed by the waterproof breathable film.

In some embodiments of the present application, a first barrier is disposed on the entrance flow channel, and a second barrier is disposed on the exit flow channel.

In some embodiments of the present application, four amplification chambers are provided, and are arranged circumferentially at equal intervals around the center of the central chamber.

In some embodiments of the present application, one amplification chamber and the corresponding entry flow channel and exit flow channel thereof form a wing-shaped amplification unit, and four amplification units are symmetrically arranged to form a butterfly-wing-shaped amplification unit group.

In some embodiments of the present application, the gas-containing areas are at least four and are respectively located between every two adjacent amplification units.

In some embodiments of the present application, a transition gas zone is further disposed between the gas-containing zones; the transition gas area is communicated with the gas containing area adjacent to the transition gas area through a gas distributing passage.

In some embodiments of the present application, the valve is a thimble valve structure.

In some embodiments of the present application, the valve comprises a valve cavity, an elastomeric gasket, and at least one protrusion;

the side surface of the valve cavity is provided with a first flow passage hole, and the bottom of the valve cavity is provided with a second flow passage hole; the elastic cushion is positioned above the second flow passage hole; the convex part is arranged on the surface of the elastic cushion facing the valve cavity, and/or is arranged on the surface of the valve cavity at a position corresponding to the elastic cushion, so that a gap is formed between the elastic cushion and the second flow passage hole.

In some embodiments of the present application, the second flow passage hole is provided at a center of the valve cavity; the convex part is arranged on the surface of the valve cavity and is positioned on the connecting line of the first flow passage hole and the second flow passage hole.

In some embodiments, the first flow passage hole is disposed at an upper portion of a side surface of the valve chamber, and a corresponding lower portion of the first flow passage hole is inclined downward from outside to inside.

In some embodiments of the present application, the reagent pack set comprises a housing, a sealing membrane, and an attachment;

the shell is provided with at least four liquid containing cavities with openings facing the front side, corresponding test solutions are preset in the liquid containing cavities, and the sealing films are attached to the front side of the shell to seal the liquid containing cavities, so that corresponding reagent pack units are formed; one side of the attachment is used for being connected with the front face of the shell, the other side of the attachment is used for being connected with the surface of the cassette board, and a hollow part is formed in the position, corresponding to the liquid containing cavity, of the attachment.

In some embodiments of the present application, the first air flow channel, the second air flow channel, the third air flow channel and the fourth air flow channel each include an upper air channel, a test solution cavity and a lower flow channel; one end of the upper air passage is provided with the vent, and the other end of the upper air passage is communicated with the upper part of the test solution cavity; one end of the lower flow channel is communicated with the lower part of the test solution cavity, and the other end of the lower flow channel is connected into the main flow channel; the reagent pack unit is broken and then the built-in test solution flows into the reagent chamber and the lower runner in sequence.

In some embodiments of the present application, a bale breaker is further included; the bag breaking piece is arranged at a position, with a preset distance, in the test solution cavity and the sealing film, and the bag breaking piece breaks the sealing film when being subjected to preset pressure.

In some embodiments of the present application, the bale breaking member is a thermoplastic elastomer;

and the edge of the test solution cavity is provided with injection molding grooves, all the injection molding grooves of the test solution cavity are communicated with each other to form an injection molding runner, and the thermoplastic elastic glue is formed in the injection molding runner by thermoplastic elastic materials.

In some embodiments of the present application, the middle portion of the test solution chamber is further provided with a separation strip, the test solution chamber is divided into an upper chamber and a lower chamber which are communicated with each other, and the separation strip is provided with a separation injection molding groove communicated with the injection molding groove on the surface.

In some embodiments of the present application, the arrangement rule of the liquid containing cavities on the shell is as follows: the liquid containing cavities are arranged in a plurality of rows from top to bottom, the liquid containing cavities in each row are arranged at equal intervals, and the liquid containing cavities in even rows correspond to gaps between every two adjacent liquid containing cavities in odd rows respectively.

In some embodiments of the present application, at least one flow guide group is arranged in the liquid containing cavity, and the flow guide group comprises two flow guide plates which are symmetrically arranged on two sides of the liquid containing cavity and are inclined downwards.

In some embodiments of the present application, the closure includes a sample application plate and a plug cap;

the sample adding plate is provided with a sample adding port and a first fixing part; the sample adding plate is arranged at the top of the sample cavity;

the plug cover comprises a base plate, a plunger and a second fixing part; one end of the plunger is arranged on the bottom surface of the substrate, and the other end of the plunger is inserted into the sample adding port to seal the sample adding port;

one end of the second fixing part is arranged on the bottom surface or the side surface of the substrate, and the other end of the second fixing part is connected with the first fixing part.

In some embodiments of the present application, the first fixing portion is a fixing hole, and the second fixing portion is a fixing strip; one end of the fixing strip is connected with the side face of the base plate through a bendable piece.

In some embodiments of the present application, the end of the fixing strip for inserting into the fixing hole is provided with a locking member, and the locking member makes the fixing strip inserted into the fixing hole and then can not be pulled out.

In some embodiments of the present application, a sample adding tube is extended from the sample adding opening, and the plunger is in a shape of a plunger matched with the sample adding tube; the inner surface of one end of the sample adding pipe, which is far away from the sample adding plate, is provided with at least one circle of inner grooves; the outer surface of the corresponding position of the plunger is provided with an outer protruding strip matched with the inner groove; one end of the sample adding pipe close to the sample adding plate is provided with a second inclined part from outside to inside; and the corresponding position of the plunger is provided with an inclined structure matched with the second inclined part.

The application also provides a test device comprising a cartridge as claimed in any one of the preceding claims and a kit.

Compared with the prior art, the card box and the detection device provided by the embodiment of the invention have the beneficial effects that:

according to the card box and the detection device provided by the embodiment of the invention, each reaction cavity is arranged on the card box body, the reagents required by the reaction are independently sealed in the reagent pack group attached to the card box body, and the main flow channel, the airflow channel and the valve which are arranged on the card box body are controlled by the matched instrument, so that the reagent pack is selectively communicated with the reaction cavities, the automatic treatment of nucleic acid extraction, amplification and detection can be realized, only a few manual operation steps are needed, the operation is simple, the safety and convenience are realized, the sealing performance is strong, and the detection result is more accurate.

Drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings needed for the embodiments or the prior art descriptions will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and it is obvious for those skilled in the art to obtain other drawings without creative efforts.

FIG. 1 is a schematic view of an exploded structure of a cartridge of an embodiment of the invention;

FIG. 2 is a schematic front view of the cassette plate;

FIG. 3 is a schematic view of the back side of the cassette plate;

FIG. 4 is a schematic view of a main flow channel of the cassette plate;

FIG. 5 is a front isometric view of the cassette plate;

FIG. 6 is a schematic front view of the cartridge with the front film removed;

FIG. 7 is a schematic view of the back side of the cartridge of FIG. 6;

FIG. 8 is a front top structural schematic view of the cartridge of FIG. 6;

FIG. 9 is a schematic side view of the cartridge of FIG. 6;

FIG. 10 is a schematic diagram of the front structure of the PCR chamber;

FIG. 11 is a schematic diagram I of the front axial side structure of the PCR chamber;

FIG. 12 is a schematic diagram II of the front axial side structure of the PCR chamber;

FIG. 13 is a schematic view of the structure of a central chamber and one amplification chamber;

FIG. 14 is an exploded view of the spike valve structure;

FIG. 15 is a schematic structural view of a spike valve structure;

FIG. 16 is a schematic front view of a spike valve structure;

FIG. 17 is a schematic view of a rear side of the spike valve structure;

FIG. 18 isbase:Sub>A cross-sectional view taken at A-A of FIG. 16;

FIG. 19 is a schematic axial side view of the housing;

FIG. 20 is a front view of the housing;

FIG. 21 is a schematic top view of the housing;

FIG. 22 is a schematic view of the structure of the gas flow passage area of FIG. 4;

FIG. 23 is a schematic structural view of the gas flow passage region of FIG. 5;

FIG. 24 is a schematic isometric view of the flow channel region;

FIG. 25 is a schematic view of the mating structure of the cap with the sample addition chamber;

FIG. 26 is a schematic view of the structure of a loading port, a loading tube and a sample loading chamber;

FIG. 27 is a first schematic view of the construction of the closure;

FIG. 28 is a second schematic structural view of the closure;

FIG. 29 is a front view of the closure;

in the figure:

1. a card box body; 11. a cassette plate; 12. a front film; 13. a rear film;

101. a sample addition chamber; 102. an internal reference freeze-dried bead cavity; 103. a sample processing chamber; 104. a magnetic bead storage chamber; 105. a waste fluid chamber; 106. a magnetic bead capture chamber; 107. a mixing chamber; 108. a blocking chamber; 109. a PCR chamber; 110. siphon bent pipe; 111. a sample addition port;

2. a reagent pack group; 21. a housing; 22. sealing the film; 23. attaching an accessory; 24. a limiting strip; 25. a baffle; 201. a first liquid containing chamber; 202. a second liquid containing cavity; 203. a third liquid containing cavity; 204. a fourth liquid containing cavity; 205. a fifth liquid containing cavity; 121. a lysis solution bag; 122. combining the liquid bag; 123. a first cleaning solution bag; 124 an eluate package; 125. a second cleaning liquid bag;

3. sealing the cover; 31. adding a sample plate; 311. a fixing hole; 312. a sample adding pipe; 32. a plug cover; 321. a substrate; 322. a plunger; 323. a fixing strip; 324. a bendable member; 325. a locking member; 326. introducing a strip; 327. an outer protruding strip;

a1, a first air flow channel; a2, a second air flow channel; a3, a third air flow channel; a4, a fourth gas flow channel; a5, a fifth air channel; a6, a sixth air passage; a7, a seventh airway; a8, an eighth air passage;

131. an upper airway; 132. a test solution chamber; 133. a lower runner; 134. injection molding a groove; 135. a dividing strip; 136. separating injection molding grooves;

141. a first branch; 142. a second branch circuit; 143. a third branch; 144. a fourth branch; 145. a fifth branch; 146. a sixth branch; 147. a seventh branch; 148. an eighth branch; n1, a positive groove; n2, back grooves; n3, a communication hole;

b1, a first valve; b2, a second valve; b3, a third valve; b4, a fourth valve; b5, a fifth valve; b6, a sixth valve; b7, a seventh valve; b8, an eighth valve; b9, a ninth valve;

151. a valve cavity; 152. an elastic pad; 153. a convex portion; 154. a first flow channel hole; 155. a second flow channel hole;

161. a central lumen; 162. an amplification chamber; 163. a zone flow channel; 1631. an inlet flow channel; 1632. an outlet flow channel; 164. a central runner channel; 165. rong Qiou; 1651. a first gas containment zone; 1652. a second gas containment zone; 166. a waterproof breathable film; 167. a zone gas duct; 168. air dividing; 169. a transition gas zone; m1, first barrier; m2, second blocking;

4. and (4) a bag breaking piece.

Detailed Description

The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.

In the description of the present invention, it is to be understood that the terms "upper", "lower", "left", "right", "top", "bottom", and the like, indicate orientations or positional relationships based on those shown in the drawings, and are only for convenience in describing the present invention and simplifying the description, but do not indicate or imply that the device or element referred to must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present invention.

It will be understood that the terms "length," "width," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like, as used herein, refer to an orientation or positional relationship indicated in the drawings, which is solely for the purpose of facilitating the description and simplifying the description, and do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed and operated in a particular orientation, and is therefore not to be construed as limiting the invention.

Furthermore, the terms "first", "second" and "first" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implicitly indicating the number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.

As shown in fig. 1 to 29, a cartridge according to a preferred embodiment of the present invention comprises a cartridge body 1, a reagent pack group 2, and a cover 3.

Specifically, the card box body 1 is provided with a main flow channel, a valve group, a reaction cavity and an airflow channel. The valve set comprises a plurality of valves arranged at the preset position of the main flow channel, so that one ends of the reaction chamber and the air flow channel are selectively communicated with the main flow channel, and the other end of the air flow channel is provided with an air vent for connecting an air pump.

The reaction chamber comprises a sample chamber, a magnetic bead storage chamber 104, a magnetic bead capture chamber 106, a waste liquid chamber 105, a mixing chamber 107 and a PCR chamber 109. The sample cavity is respectively communicated with the first ends of the magnetic bead storage cavity 104 and the magnetic bead capture cavity 106, the first end of the magnetic bead capture cavity 106 is also communicated with the mixing cavity 107, the opposite second end of the mixing cavity is communicated with the waste liquid cavity 105, and the mixing cavity 107 is communicated with the PCR cavity 109. The sample chamber has a sample addition port 111, and the cover 3 is used to seal the sample addition port 111. Magnetic particles, such as freeze-dried magnetic beads, are pre-disposed in the magnetic bead storage chamber 104. Freeze-dried beads are pre-placed in the mixing chamber 107.

The reagent pack group 2 is attached to the surface of the cartridge body 1, and includes at least four independently sealed reagent pack units, where the four reagent pack units are a lysis solution pack 121, a binding solution pack 122, a first cleaning solution pack 123 and an eluent pack 124, respectively, and may further preferably include a second cleaning solution pack 125 (in fig. 2 and 3, the reference numerals are only schematic of the positions of the reagent pack units, and do not indicate complete reagent pack units).

The gas flow channels include at least a first gas flow channel A1, a second gas flow channel A2, a third gas flow channel A3, and a fourth gas flow channel A4 corresponding to the lysis pad 121, the binding pad 122, the first wash pad 123, and the eluent pad 124, respectively, and may preferably further include a fifth gas flow channel A5 corresponding to the second wash pad 125.

The lysis solution bag 121 is broken and then flows into the first gas flow channel A1, and the first gas flow channel A1 may be communicated with the magnetic bead storage chamber 104 or with the sample chamber.

The binding liquid packet 122 is broken and flows into the second flow channel A2, and the second flow channel A2 can be communicated with the magnetic bead storage chamber 104 or the sample chamber.

The above steps are mainly for mixing the lysis solution, the magnetic beads (magnetic particles) in the magnetic bead storage chamber 104, and the binding solution in the sample chamber. When the first gas flow channel A1 is communicated with the magnetic bead storage cavity 104, the lysis solution carries the magnetic beads to the sample cavity, and at this time, the binding solution directly flows into the sample cavity, so that the second gas flow channel A2 is communicated with the sample cavity. When the first gas flow channel A1 is directly connected to the sample chamber, i.e. the binding solution carries the magnetic beads to the sample chamber, the second gas flow channel A2 needs to be connected to the magnetic bead storage chamber 104.

The first cleaning solution bag 123 is broken and flows into the third flow channel A3, and the third flow channel A3 may be communicated with the first end of the sample chamber or the magnetic bead capture chamber 106. This step is mainly to wash the magnetic beads, and to make the washing solution pass through the magnetic bead capture chamber 106 to wash the magnetic beads, and then inject the washing solution into the waste liquid chamber 105. Therefore, the third flow channel A3 may be directly communicated with the first end of the magnetic bead capture cavity 106 for magnetic bead cleaning, and may also be communicated with the sample cavity, so that the cleaning solution flows through the sample cavity first and then flows into the magnetic bead capture cavity 106 for magnetic bead cleaning.

The second cleaning solution bag 125 is broken and flows into the fifth flow channel A5, and the fifth flow channel A5 may be communicated with the first end of the sample chamber or the magnetic bead capture chamber 106. The step is mainly to perform a second washing on the magnetic beads, and the principle and the steps are the same as those of the first washing, which are not described herein again. The second washing of the magnetic beads is a preferred embodiment and is not an essential step.

After the eluent packet 124 is broken, the eluent flows into the fourth gas flow channel A4, and the fourth gas flow channel A4 is communicated with the second end of the magnetic bead capture cavity 106. This step is mainly to elute the nucleic acid molecules adsorbed on the surface of the magnetic beads, and then push the eluent into the mixing chamber 107 to mix the eluent with the lyophilized beads in the mixing chamber 107.

After the eluent and the freeze-dried beads are mixed in the mixing cavity 107, the mixture is pushed to the PCR cavity 109 for amplification reaction.

In the above steps, the flow of liquid is controlled by the valve set and the air pump.

According to the above process steps, the present application proposes a preferred embodiment of the cartridge as follows:

referring to fig. 1-4, the cartridge comprises a cartridge body 1, a reagent pack set 2 and a cover 3, wherein the cartridge body 1 comprises a cartridge plate 11, a front membrane 12 and a rear membrane 13. The front film 12 and the rear film 13 are respectively attached to the front and the back of the cassette board 11 for sealing the openings, the slots and the hollowed-out portions of the cavities on the front and the back of the cassette board 11. In the following description, the descriptions relating to the left, right, up, and down orientations are based on the front view of the cartridge board 11.

In this embodiment, the sample chamber preferably includes a sample addition chamber 101, a reference freeze-dried bead chamber 102, a sample processing chamber 103, and a siphon trap 110, which are disposed at predetermined positions in the middle of the front surface of the cartridge plate 11. The sample addition chamber 101 has a sample addition port 111 at the top, a sample to be tested is injected into the cartridge from the sample addition port 111, and the cover 3 is used for closing the sample addition port 111. The internal reference freeze-dried bead cavity 102 is located below the sample adding cavity 101, the top of the internal reference freeze-dried bead cavity 102 is communicated with the bottom of the sample adding cavity 101, and the bottom of the internal reference freeze-dried bead cavity 102 is communicated with one end of the siphon elbow 110. The sample processing chamber 103 is located at the left side of the sample adding chamber 101, the other end of the siphon bend 110 is communicated with the top of the sample processing chamber 103 at the position which is more right than the top, a sixth air passage A6 is arranged at the top of the left side of the sample processing chamber 103, and the other end of the sixth air passage A6 is provided with an air vent (in the drawings, for the convenience of understanding and marking, a mark line is marked at the air vent of the air passage in the figures of the application, for example, the mark line of A6 is marked at the air vent of the sixth air passage A6 in the figures, and other air passages are the same. The internal reference freeze-dried beads are preset in the internal reference freeze-dried bead cavity 102 and serve as internal reference samples, and the internal reference freeze-dried beads have the function of monitoring the whole card box process. If the internal reference freeze-dried bead sample can have a fluorescence reaction, and the sample to be detected does not have a fluorescence reaction, the problem of the sample to be detected is indicated.

The magnetic bead storage cavity 104 is located on the left side of the sample processing cavity 103, the mixing cavity 107 is located below the siphon bent pipe 110, a seventh air passage A7 is arranged at the central line of the top of the mixing cavity 107, and an air vent is formed in the other end of the seventh air passage A7. Waste liquid chamber 105 is located the right side that the chamber 101 was added to the sample, and waste liquid chamber 105's top is equipped with eighth air flue A8, and the other end of eighth air flue A8 has the blow vent, can regard as the exhaust hole. The PCR chamber 109 is provided at the right side of the front face of the cassette plate 11.

In some embodiments of the present application, a blocking chamber 108 is further disposed between the mixing chamber 107 and the PCR chamber 109. The blocking chamber 108 is located below the sample addition chamber 101 and is connected to a communication branch (main channel) between the mixing chamber 107 and the PCR chamber 109. In some embodiments of the present application, the inner top of the blocking cavity 108 is provided with a solid wax, which melts when heated to a predetermined temperature, and the wax seals the main flow channel, thereby blocking the communication between the mixing cavity 107 and the PCR cavity 109, so as to seal the PCR cavity 109 after completing the amplification reaction, and prevent the amplification product in the PCR cavity 109 from leaking out of the cartridge through the flow channel to pollute the instrument or the environment.

The reagent pack group 2 is attached to the back of the left upper side of the cassette board 11 and includes five independent sealed reagent pack units, and the five reagent pack units are a lysis solution pack 121, a binding solution pack 122, a first cleaning solution pack 123, an elution solution pack 124 and a second cleaning solution pack 125 (in the figure, the reference numerals are only schematic for the positions of the reagent pack units, and do not represent the complete reagent pack units).

Referring to fig. 4, the main flow path on the cassette board 11 includes a first branch 141, a second branch 142, a third branch 143, a fourth branch 144, a fifth branch 145, a sixth branch 146, a seventh branch 147, and an eighth branch 148.

One end of the first branch 141 is connected to the liquid outlet end of the first air flow passage A1, the other end of the first branch 141 is sequentially communicated with the magnetic bead storage chamber 104, the first valve B1 and the sample processing chamber 103.

One end of the second branch 142 is connected to the liquid outlet end of the second flow channel A2, and the other end of the second branch 142 is connected to the first branch 141 after being connected to the second valve B2, where the access point is denoted as C1.

One end of the third branch 143 is connected to the liquid outlet end of the third flow channel A3, and the other end of the third branch 143 is connected to the first branch 141 after being sequentially connected to the fourth valve B4 and the fifth valve B5, where the access point is denoted as C2.

One end of the fourth branch 144 is connected to the liquid outlet end of the fourth gas flow channel A4, and the other end of the fourth branch 144 is connected to the seventh valve B7 and one end (second end) of the magnetic bead capturing cavity 106 near the left side of the cartridge board 11.

One end of the fifth branch 145 is communicated with the liquid outlet end of the fifth flow channel A5, the other end of the fifth branch 145 is connected to the third branch 143, and the access point C3 is located between the fourth valve B4 and the fifth valve B5.

One end of the sixth branch 146 is communicated with the bottom of the sample processing chamber 103 (or connected to a section of the first branch 141 close to the sample processing chamber 103), and the other end of the sixth branch 146 is communicated with the third valve B3 and then communicated with one end (first end) of the magnetic bead capture chamber 106 close to the right side of the cassette plate 11.

One end of the seventh branch 147 is connected to the upper portion of the waste liquid chamber 105, and the other end of the seventh branch 147 is connected to the sixth valve B6 and one end (second end) of the magnetic bead capturing chamber 106 near the left side of the cartridge plate 11.

One end of the eighth branch 148 is communicated with one end (first end) of the magnetic bead capturing cavity 106 close to the right side of the cassette plate 11, and the other end of the eighth branch 148 is sequentially communicated with an eighth valve B8, a mixing cavity 107, a blocking cavity 108, a ninth valve B9 and a PCR cavity 109.

In the above embodiment, referring to fig. 1 to 9, the sample adding chamber 101, the reference freeze-dried bead chamber 102, the sample processing chamber 103, the waste liquid chamber 105, the mixing chamber 107, and the blocking chamber 108 are cavities formed by protruding the front surface of the cartridge board 11 toward the back surface thereof, and the cavities are sealed when the front film 12 is attached to the front surface of the cartridge board 11 to form the above reaction chambers. In some embodiments, the sections of the sample addition chamber 101 and the sample processing chamber 103 near the bottom are inclined toward the front surface of the cartridge plate 11 to facilitate the liquid outflow. A groove is cut in the front face of the cartridge plate 11 and sealed by the front membrane 12 to form a siphon trap 110. Referring to fig. 7, a hollow hole is formed in the cartridge plate 11, a protrusion 1041 slightly protruding out of the back surface of the cartridge plate 11 is disposed at a position around the edge of the hollow hole on the back surface of the cartridge plate 11, the protrusion 1041 is provided with a flow channel to communicate with the first valve B1, and the front surface of the hollow hole is sealed by the front film 12 to form the magnetic bead storage chamber 104.

In the above embodiments, the main flow channel includes the first branch 141, the second branch 142, the third branch 143, the fourth branch 144, the fifth branch 145, the sixth branch 146, the seventh branch 147 and the eighth branch 148, which are all formed by grooves formed on one surface (front surface or back surface) of the cassette board 11. In some embodiments, the present application provides a positive and negative trench structure, and the branch can be composed of at least one positive and negative trench structure. Referring to fig. 10, taking the eighth branch 148 as an example, the front-back groove structure includes a front groove N1, a back groove N2, and a communication hole N3. The positive groove N1 is a groove (solid line segment in the figure) formed on the front surface of the card box plate 11, the back groove N2 is a groove (dotted line segment in the figure) formed on the back surface of the card box plate 11, and the communication hole N3 is formed in the intersection of the ports of the positive groove N1 and the back groove N2 and used for communicating the ports of the positive groove N1 and the back groove N2. The front film 12 seals the front groove N1, the rear film 13 seals the back groove N2, and the communication hole N3 communicates the front groove N1 and the back groove N2, thereby forming a flow passage.

In the above-described embodiment of the present invention, referring to FIGS. 10-12, PCR chamber 109 includes a central chamber 161, an amplification chamber 162, a zone flow channel 163, a central flow channel groove 164, a gas containing zone 165, a water-proof gas-permeable membrane 166, a zone gas channel 167, a gas dividing channel 168, and a transition gas zone 169.

The central flow channel groove 164 is formed on one surface (front surface) of the central chamber 161, the flow channel (i.e., the back recess N2 section of the eighth branch 148) is formed on the other surface (back surface) of the central chamber 161 opposite to the central chamber, the eighth branch 148 is communicated with one end of the central flow channel groove 164, and the other end of the central flow channel groove 164 is communicated with the block flow channel 163. A waterproof, breathable membrane 166 is attached to the central cavity 161 to seal the central runner channel 164. The regional air channel 167 is arranged at the edge of the sealing area of the waterproof breathable film 166, and the regional air channel 167 is communicated with the air containing region 165. Referring to fig. 12, the height of the central channel 164 is about half of the height of the central cavity 161. When the waterproof membrane 166 is attached to the central cavity 161, the upper half of the central cavity 161 is not sealed by the waterproof membrane 166, and the gas channel 167 is actually a channel between the upper half of the central cavity 161 and the gas containment region 165. In this embodiment, a waterproof air-permeable film 166 is disposed on the opposite surface (front surface) of the liquid inlet end (the eighth branch 148 disposed on the back surface), and the sealed edge thereof is communicated with Rong Qiou. When the mixed liquid in the mixing chamber 107 enters the central channel groove 164 of the central chamber 161, the carried air bubbles will be released from the waterproof and air-permeable membrane 166 and enter the air-holding region 165, so that the mixed liquid entering the amplification chamber 162 does not carry air bubbles, thereby facilitating the amplification step.

In some embodiments of the present application, referring to fig. 10 and fig. 13, one amplification cavity 162 is correspondingly provided with two region runners 163, and the two region runners 163 are an entrance runner 1631 and an exit runner 1632, respectively. One end of the entrance flow channel 1631 communicates with the central flow channel 164, and the other end of the entrance flow channel 1631 communicates with one end of the amplification chamber 162. One end of the exit flow channel 1632 is communicated with the other end of the amplification chamber 162, and the port of the other end of the exit flow channel 1632 is located in the central chamber 161 and sealed by the waterproof and breathable membrane 166. The mixed liquid in the central channel groove 164 enters the amplification chamber 162 through the inlet channel 1631, when the amplification chamber 162 is filled with the mixed liquid, the excess mixed liquid flows into the outlet channel 1632, and when the outlet channel 1632 is filled, the mixed liquid cannot pass through the waterproof air-permeable membrane 166, so that the mixed liquid stops entering the inlet channel 1631.

In some embodiments of the present application, a first barrier M1 is disposed on the inlet flow channel 1631, and a second barrier M2 is disposed on the outlet flow channel 1632. In practice, the first barrier M1 and the second barrier M2 are a flow path interrupting structure of a section of the unwelded front film 12. Referring specifically to fig. 13, when the front film 12 is welded on the surface of the cassette board 11 by laser, the welding line is about 0.1mm away from the outside of the flow channel, i.e., the area outside the dotted line strip circle X in the figure is the welding area. Since the first barrier M1 and the second barrier M2 are non-welded regions, and the front film 12 is only attached to the surface of the cartridge board 11, when the flow rate or pressure of the mixed liquid is high, the front film 12 at the barrier is pressed to arch the front film 12 outwards, and the mixed liquid can still flow through the barrier (as shown by the arrow in the figure). The first barrier M1 and the second barrier M2 can block flow to some extent, but do not completely block flow, so that the mixed liquid can flow among the inlet flow channel 1631, the amplification chamber 162 and the outlet flow channel 1632. When the amplification chamber 162 is filled, the flow of the inlet flow channel 1631 and the outlet flow channel 1632 needs to be completely blocked before the temperature rise and drop cycle, at this time, the waterproof breathable membrane 166 is a first barrier, and the top pressure rod of the matching instrument is used to press the front membrane 12 at the first barrier M1 and the second barrier M2 to form a second barrier, thereby ensuring that the flow of the inlet flow channel 1631 and the outlet flow channel 1632 is completely blocked.

In some embodiments, four amplification chambers 162 are provided, and are arranged circumferentially at equal intervals around the center of the central chamber 161. One amplification cavity 162 and the corresponding entry flow channel 1631 and exit flow channel 1632 form a wing-shaped amplification unit, and the four amplification units are symmetrically arranged to form a wing-shaped amplification unit group. In some embodiments of the present application, the number of Rong Qiou is at least four, and each of the at least four amplification units is located between every two adjacent amplification units. A transition air area 169 is also arranged between every two adjacent air containing areas 165, and the transition air area 169 is communicated with the air containing area 165 adjacent to the transition air area through an air distributing channel 168. The central cavity 161 and the waterproof breathable membrane 166 are both circular, the center of the central cavity 161 is taken as the circle center, the position of the flow channel is 0 degree, and the four amplification cavities 162 are respectively positioned at the positions of 45 degrees, 135 degrees, 225 degrees and 315 degrees on the circle. Specifically, the air-containing region 165 includes a first air-containing region 1651 and a second air-containing region 1652. The first air-containing area 1651 is provided with two air-containing areas, which are respectively located at the axes of 90 ° and 270 °, and the shape of each air-containing area is similar to a triangle corresponding to the fin-shaped gap, one angle of each air-containing area is communicated with the central cavity 161 through the air-distributing channel 168, and the other two angles of each air-containing area are respectively communicated with one end of the transition air area 169 through the air-distributing channel 168. The second air-containing area 1652 is provided with four air-distributing channels 168, two of which are located on two sides of the flow channel, and the other two air-distributing channels are symmetrically located on two opposite sides of the flow channel, and the air-distributing channels 168 and the air-distributing area 167 are communicated with the central cavity 161. The other end of transition gas zone 169 communicates with second gas containing zone 1652 via gas dividing duct 168. Referring to fig. 11-12, the air containing region 165 and the transition air region 169 are hollow regions, the air dividing duct 168 and the air dividing duct 167 are grooves formed on the card case body 1, and the arrangement form of the region flow passage 163 is a positive and negative groove structure form. After the mixed liquid in the mixing chamber 107 flows into each amplification chamber 162, the optical module of the matched instrument is positioned at the corresponding position of the front PCR chamber 109 of the card box, the thermal module is positioned at the corresponding position of the back PCR chamber 109 of the card box, and the optical module and the thermal module clamp the card box from the two sides of the card box. During amplification, the thermal module cyclically heats and cools the amplification chamber 162. After amplification is complete, it is desirable to de-excite the amplified reactants in the amplification chamber 162 with light of different wavelengths to detect the concentration of the amplified reactants in the amplification chamber 162. Specifically, the optical module includes four small modules, namely a red module, a green module, a yellow module and a blue module, the four small modules are respectively disposed on the four amplification chambers 162 to detect different types of fluorescence after reaction of reagents in the amplification chambers 162, and each amplification chamber 162 needs to receive detection of the four small modules in turn. Because the four amplification cavities 162 are designed on one circumference, the optical module can correspondingly detect different amplification cavities 162 by the four small modules in turn only by rotating, and the optical module does not need to move an extra stroke, thereby effectively improving the detection efficiency.

In the above embodiment, the valves included in the valve group include the first valve B1, the second valve B2, the third valve B3, the fourth valve B4, the fifth valve B5, the sixth valve B6, the seventh valve B7, the eighth valve B8, and the ninth valve B9. The valve can adopt any valve body structure which is suitable for a card box structure in the prior art and can realize the liquid flow control through being matched with equipment instruments, wherein, a thimble valve structure is preferably adopted.

In some embodiments, the present application provides a preferred spike valve structure.

Referring to FIGS. 14-18, the present application contemplates a spike valve structure comprising a valve cavity 151, a resilient pad 152, and at least one protrusion 153. The valve chamber 151 has a first flow passage hole 154 formed in a side surface thereof and a second flow passage hole 155 formed in a bottom surface thereof. The elastic pad 152 is located above the second flow path hole 155. The protrusion 153 is provided on a surface of the elastic pad 152 facing the valve chamber 151 and/or is provided on a surface of the valve chamber 151 at a position corresponding to the elastic pad 152 such that a gap is provided between the elastic pad 152 and the second flow path hole 155. Preferably, the second flow passage hole 155 is provided at the center of the valve chamber 151. One protrusion 153 is provided, and the protrusion 153 is provided on the surface of the valve chamber 151 on the line connecting the first flow channel hole 154 and the second flow channel hole 155. In some embodiments of the present application, the shape of the protrusion 153 is a dome shape. In some embodiments, the valve cavity 151 is circular, and the elastic pad 152 is also circular, and the diameter of the elastic pad 152 is slightly smaller than the diameter of the valve cavity 151, and the elastic pad 152 is preferably a silicone pad. In some embodiments, the side of the valve cavity 151 is sloped downward from the outside to the inside. In some embodiments, the first flow passage holes 154 are disposed at the upper side of the valve chamber 151, and the corresponding lower side is inclined downward from the outside to the inside to facilitate the liquid flowing into the valve chamber 151.

In the structure of the present invention, due to the protrusion 153, when the elastic pad 152 is not subjected to an external force, a gap is formed between the elastic pad 152 and the second flow channel hole 155, so that the liquid can flow into the valve chamber 151 from the first flow channel hole 154 and then flow into the second flow channel hole 155, and then flow out of the structure of the top needle valve along the flow channel. When the elastic pad 152 is subjected to a predetermined pressure, it is deformed, and the elastic pad 152 blocks the second flow passage hole 155, so that the liquid cannot flow into the second flow passage hole 155, i.e. the structure of the needle valve functions to block the flow of the liquid. Because the elastic pad 152 has elasticity, when the external force is removed, the elastic pad 152 is restored to the original state, and the elastic pad 152 and the second flow passage hole 155 have a gap therebetween, so that the liquid can flow through the needle valve structure.

The valve cavity 151 of the thimble valve structure provided by the application can be directly opened on the front surface of the card box plate 11, and the flow channel with the positive and negative groove structure is adopted in the above embodiment, so that the first flow channel hole 154 is communicated with the positive groove N1 arranged on the front surface of the card box plate 11, the second flow channel hole 155 is communicated with the back groove N2 arranged on the back surface of the card box plate 11, and the thimble valve structure can be regarded as a communicating hole N3 with a controllable flow blocking function, and the thimble valve structure is simple in structure and convenient to control and reliable.

In the above embodiment, referring to fig. 1, the reagent pack set 2 is attached to the back surface of the left upper side of the cassette board 11, and the reagent pack set 2 includes the housing 21, the sealing film 22, and the attachment 23.

Referring to fig. 19-21, the housing 21 is provided with five liquid-containing chambers with their openings facing the front, each of which includes: a first liquid holding cavity 201 with a lysis solution preset inside, a second liquid holding cavity 202 with a binding solution preset inside, a third liquid holding cavity 203 with a cleaning solution preset inside, a fourth liquid holding cavity 204 with an eluent preset inside, and a fifth liquid holding cavity 205 with a secondary cleaning solution preset inside. The sealing film 22 is attached to the front surface of the housing 21 to seal the liquid containing cavity, thereby forming five reagent pack units, namely a lysis solution pack 121, a binding solution pack 122, a first cleaning solution pack 123, an eluent pack 124 and a second cleaning solution pack 125. One surface of the attachment 23 is used for connecting the front surface of the shell 21, the other surface of the attachment 23 is used for connecting the back surface of the cassette board 11, and a hollow part is formed at a position corresponding to the liquid containing cavity on the attachment 23, namely the attachment 23 does not shield the sealing film 22.

In some embodiments of the present application, referring to fig. 22, each of the first air flow channel A1, the second air flow channel A2, the third air flow channel A3, the fourth air flow channel A4, and the fifth air flow channel A5 includes an upper air channel 131, a test solution cavity 132, and a lower air channel 133, one end of the upper air channel 131 has an air vent, and the other end thereof communicates with an upper portion of the test solution cavity 132. One end of the lower channel 133 communicates with the lower portion of the sample chamber 132, and the other end thereof is connected to the main channel. The test solution chamber 132 corresponds to the solution containing chamber, and the reagent pack unit is broken and the built-in test solution flows into the test solution chamber 132 and the lower flow channel 133 in sequence. Specifically, the lower flow channel 133 of the first flow channel A1 communicates with one end of the first branch 141 of the main flow channel, the lower flow channel 133 of the second flow channel A2 communicates with one end of the second branch 142 of the main flow channel, the lower flow channel 133 of the third flow channel A3 communicates with one end of the third branch 1431 of the main flow channel, the lower flow channel 133 of the fourth flow channel A4 communicates with one end of the fourth branch 144 of the main flow channel, and the lower flow channel 133 of the fifth flow channel A5 communicates with one end of the fifth branch 145 of the main flow channel.

In the above embodiment, referring to fig. 22-24, the position of the cassette plate 11 corresponding to the liquid containing cavity is hollowed out to form the liquid testing cavity 132. The upper air passage 131 and the lower air passage 133 are formed by alternately grooving the front and the back of the cassette plate 11, i.e., the upper air passage 131 and the lower air passage 133 can adopt a positive and negative groove structure.

In some embodiments, the present application further comprises a bale breaker 4. The bag breaking piece 4 is arranged in the test solution cavity 132 and has a preset distance with the sealing membrane 22, and the bag breaking piece 4 breaks the sealing membrane 22 when being subjected to a preset pressure.

In some embodiments of the present application, the bale breaker 4 is a thermoplastic elastomer. Specifically, referring to FIGS. 22-24, the edges of the test solution chamber 132 are provided with injection molding grooves 134. In some embodiments of the present application, a partition bar 135 is further disposed at a middle portion of the test solution cavity 132 to partition the test solution cavity 132 into an upper cavity and a lower cavity which are communicated with each other, and a partition injection molding groove 136 communicated with the injection molding groove 134 is disposed on a surface of the partition bar 135. The injection molding grooves 134 and the partition injection molding grooves 136 of all the test solution chambers 132 are communicated with each other to form an injection molding runner, and the thermoplastic elastomer is molded into a block of the thermoplastic elastomer in the injection molding runner, as shown in fig. 1 and 6. Correspondingly, referring to fig. 1, the front film 12 is hollowed out at a position corresponding to the bag breaking member 4, so that the bag breaking member 4 is pressed by a pressing module of a matching instrument.

During the actual use, the thermoplastic elastic glue is extruded to the pressing module of supporting instrument, utilizes the good ductility of thermoplastic elastic glue to extrude and break sealing membrane 22, lets the test solution of holding the liquid intracavity flow out. In this embodiment, utilize the edge design of test solution chamber 132 to mould plastics the runner so that the shaping is glued to thermoplasticity elasticity, and the broken package 4 that forms like this can laminate in test solution chamber 132 department, effectively avoids broken package 4 to shift. In addition, the middle part of the test solution cavity 132 is provided with the separating strip 135 with the separating injection molding groove 136, so that the thermoplastic elastic adhesive can be supported, and a matched instrument can extrude and break the package from the upper cavity and the lower cavity at the same time, thereby improving the package breaking efficiency.

In some embodiments of the present application, the arrangement rule of the liquid containing cavities on the casing 21 is: a plurality of rows of liquid containing cavities are arranged from top to bottom, the liquid containing cavities in each row are arranged at equal intervals, and the liquid containing cavities in even rows correspond to the gaps between every two adjacent liquid containing cavities in odd rows respectively. For example, in the present embodiment, five liquid containing chambers are provided, and the arrangement of the first liquid containing chamber 201, the second liquid containing chamber 202, the third liquid containing chamber 203, the fourth liquid containing chamber 204, and the fifth liquid containing chamber 205 is as shown in fig. 20. The arrangement of the liquid containing cavities facilitates the better arrangement of the flow channels, and as shown in fig. 22, the upper air channel 131 of the second flow channel A2 passes through the test solution cavities 132 corresponding to the first flow channel A1 and the fifth flow channel A5, and is communicated with the test solution cavities 132 corresponding to the second flow channel A2.

In some embodiments of the present application, referring to fig. 18-19, at least one flow guide set is disposed in the liquid containing cavity, and the flow guide set includes two flow guide plates 25 symmetrically disposed at two sides of the liquid containing cavity and inclined downward. In some embodiments of the present application, the liquid chamber has a long bar shape, and two ends of the liquid chamber form an arc shape protruding upward and downward, respectively. In some embodiments of the present application, the middle dividing line of the liquid containing cavity divides the liquid containing cavity into an upper liquid containing cavity and a lower liquid containing cavity, and the two flow guiding groups are provided, one is provided in the upper liquid containing cavity, and the other is provided in the lower liquid containing cavity. The structural design of the liquid containing cavity and the guide plate 25 are beneficial to ensuring that the test liquid in the liquid containing cavity flows out completely as much as possible after the bag is broken.

In some embodiments of the present application, the sealing film 22 may be a whole film for sealing the liquid containing cavity, or may include a plurality of single sealing films having the same number as the liquid containing cavities, where the shape of the single sealing film matches the liquid containing cavity, and one single sealing film seals one liquid containing cavity. Preferably, the sealing film 22 is an aluminum plastic film.

In some embodiments of the present application, the attachment 23 is preferably a pressure sensitive adhesive, and the shape of the attachment is consistent with the shape of the housing 21, so that the housing 21 has the pressure sensitive adhesive at the positions except for the liquid containing cavity, which is favorable for being firmly attached to the back of the cassette board 11. In some embodiments of the present application, referring to fig. 3 and 7, the back of the cassette board 11 is provided with a plurality of position-limiting strips 24 conforming to the shape of the housing 21, and the edges of the attachment 23 are located in the position-limiting strips 24 to prevent the reagent pack set 2 from shifting.

In the above embodiment, referring to fig. 25-29, the closure 3 includes a dosing plate 31 and a stopper cap 32. The sample addition plate 31 has a sample addition port 111 and a fixing hole 311 (first fixing portion). The plug cover 32 includes a base 321, a plunger 322, and a fixing bar 323 (second fixing portion). One end of the plunger 322 is disposed on the bottom surface of the base plate 321, and the other end thereof is inserted into the sample addition port 111 to seal the sample addition port 111. One end of the fixing bar 323 is provided on the bottom or side of the base 321, and the other end thereof is inserted into the fixing hole 311.

In some embodiments of the present application, one end of the fixing bar 323 is connected to the side of the base plate 321 by a bendable member 324. In some embodiments, the position of the fixing hole 311 corresponds to the position of the fixing bar 323 when the fixing bar is bent to be perpendicular to the sample feeding plate 31. When the sample addition plate 31 is positioned on the horizontal plane, that is, when the fixing strip 323 is bent to 90 °, it is just inserted into the fixing hole 311 vertically.

In some embodiments of the present invention, the end of the fixing bar 323 used for inserting into the fixing hole 311 is provided with a locking member 325, and the locking member 325 prevents the fixing bar 323 from being pulled out after being inserted into the fixing hole 311. In some embodiments of the present invention, the edge of the fixing hole 311 forms a first inclined portion from the outside to the inside, and the locker 325 is a resilient cone that is engaged with the fixing hole 311. In some embodiments, the securing hole 311 is square in shape and the locking member 325 is frustoconical in shape. In some embodiments of the present application, the front end of the locking member 325 is provided with a guiding strip 326, and the width of the guiding strip 326 is smaller than the width of the fixing hole 311, so that the guiding strip 326 plays a role of guiding the fixing strip 323 to be inserted into the fixing hole 311, thereby facilitating the operation of a technician.

In some embodiments, the sample port 111 is extended with a sample application tube 312, and the plunger 322 is shaped to match the sample application tube 312. In some embodiments of the present application, at least one circle of inner grooves is formed on an inner surface of an end of the sample adding tube 312 away from the sample adding plate 31, and an outer protrusion 327 engaged with the inner grooves is formed on an outer surface of the plunger 322 at a corresponding position. In some embodiments, the sample application tube 312 has a second inclined portion formed from the outside to the inside near one end of the sample application plate 31, and the plunger 322 is provided with an inclined structure corresponding to the second inclined portion.

In this embodiment, the sample adding plate 31 is arranged at the top of the sample adding cavity 101, the sample adding plate 31 is connected with the surface of the sample adding cavity 101, the sample adding pipe 312 extends into the sample adding cavity 101, and the sample adding pipe 312 is arranged to facilitate the sample adding of the pipette.

The closing cap 3 that this application provided makes gag 32 and sample addition port 111 department have simple connection cooperation structure (fixed strip 323 and fixed orifices 311), prevents effectively that the gag 32 from losing, avoids bringing unnecessary trouble for detection achievement. And, having carried out ingenious design to many places connection structure, made sealed more reliable, helped improving the leakproofness of card box application of sample back.

The invention also provides a detection device, which comprises the card box and a matched instrument matched with the card box. The supporting instrument mainly includes: the air pump is connected with the air port of each air passage; the positions corresponding to the valves are provided with driving modules (central rods) which are used for driving the valves to open and close; magnetic attraction modules are arranged at positions corresponding to the magnetic bead capture cavity 106 and the sample processing cavity 103 and used for providing magnetic attraction force and controlling attraction or release of magnetic beads; an ultrasonic module and a thermal cycle module are arranged at the positions corresponding to the sample processing cavity 103 and can be used for uniformly mixing and heating the sample mixed liquid; a pressing module is arranged at the position corresponding to the bag breaking piece 4 and used for providing preset pressing force and extruding the bag breaking piece 4 to break the sealing film of the reagent bag; and an amplification heating module and an optical detection module are arranged at the position corresponding to the PCR cavity 109, the amplification heating module is used for heating and cooling the PCR cavity, and the optical detection module is used for detecting the amplification condition in the PCR cavity.

The above embodiment proposes a cartridge structure, the work flow of which is as follows:

first step, loading sample: the sample of the patient to be tested is introduced into the sample addition chamber 101 from the sample addition port 111, and then the cover 3 is fastened, thereby sealing the cartridge. Because the siphon bend 110 is disposed between the sample addition chamber 101 and the sample processing chamber 103, the sample temporarily stays in the siphon bend 110 and does not directly flow into the sample processing chamber 103.

Step two, connecting a matched instrument: the cartridge is inserted into a mating instrument. Specifically, the matched instrument is provided with a slot, so that the card box can be vertically inserted into the matched instrument. The ventilation ports of the first air channel A1, the second air channel A2, the third air channel A3, the fourth air channel A4, the fifth air channel A5, the sixth air channel A6 and the seventh air channel A7 on the card box body 1 are all connected with an air pump of a matched instrument, and the connection part can be provided with an air pressure valve. Moreover, the central rods of the matching instrument are respectively aligned with the valves (the first valve B1, the second valve B2, the third valve B3, the fourth valve B4, the fifth valve B5, the sixth valve B6, the seventh valve B7, the eighth valve B8 and the ninth valve B9), and the initial states of all the central rods are the states of extruding the elastic pads so as to close all the valves, so that all the channels in the card box body 1 are in the closed state.

Step three, sample pretreatment: sample pretreatment: after sample application, the sample is mixed with the internal reference lyophilized beads in the internal reference lyophilized bead cavity, and then reaches the siphon elbow 110 to stay. After the test is started, the sample staying in the siphon bend 110 is sucked into the sample processing chamber 103 by using the air pump of the kit to suck air from the air vent of the sixth air passage A6 (the following description does not refer to the operation of the air pump of the kit, and only describes the air ventilation or air suction state of the air passage).

Step four, releasing lysate: the pressing module of the matching instrument presses the area of the bag breaking member 4 corresponding to the lysis solution bag 121, so as to puncture the sealing membrane 22 of the lysis solution bag 121, and the lysis solution flows out of the lysis solution bag 121 to the test solution cavity 132 of the first air flow channel A1 (the operation of the pressing module of the matching instrument is not mentioned, and only the state of the reagent bag is described below). At this time, since all the valves of the channel are closed, the lysate is temporarily retained in the sample chamber 132 and does not automatically flow into each chamber of the cartridge body 1. Then, the center lever corresponding to the first valve B1 is released, thereby opening the first valve B1 (the following description will not refer to the operation of the center lever of the kit, and only the open/close state of the valve will be described). Meanwhile, air is introduced from the first air flow channel A1, so that the lysate flows into the magnetic bead storage cavity 104, freeze-dried magnetic beads in the magnetic bead storage cavity 104 are uniformly mixed, and all the magnetic beads are carried to flow into the sample processing cavity 103. By continuing to introduce air from the first air flow channel A1, the lysate is sufficiently mixed with the sample by the bubbles continuously generated from the bottom of the sample processing chamber 103, and then the first valve B1 is closed. In the mixing process, the mixed solution can be heated to 85 ℃ through a thermal cycle module of a matched instrument for thermal cracking.

Step five, releasing the binding solution: the binding liquid pack 122 is punctured, and the binding liquid flows out from the binding liquid pack 122 to the sample liquid chamber 132 of the second flow channel A2. At this time, since the tip valve corresponding to the channel is in the closed state, the binding solution is temporarily retained in the sample solution chamber 132 and does not automatically flow into each chamber of the cartridge body 1. The second valve B2 is opened and air is introduced from the second flow channel A2 to push the binding solution into the sample processing chamber 103. And continuously introducing air from the second air flow channel A2, mixing the binding solution and the magnetic beads with the lysed sample by using the bubbles continuously generated from the bottom of the sample processing chamber 103, and then closing the second valve B2.

In the above steps, a mode of firstly releasing the lysis solution and mixing the lysis solution with the magnetic beads in the sample processing chamber 103, and then releasing the binding solution to the sample processing chamber 103 for mixing together is adopted. However, in the present embodiment, a manner of releasing the lysis solution into the sample processing chamber 103, releasing the binding solution to mix with the magnetic beads, and then mixing with the magnetic beads in the sample processing chamber 103 may also be adopted, and the specific ventilation and valve control in this manner are the same as in the above manner, and will not be described again here.

Sixthly, cleaning magnetic beads: in this embodiment, there are two types of magnetic bead cleaning methods.

The first magnetic bead cleaning mode is to adsorb the magnetic bead in the magnetic bead capturing cavity 106 for cleaning, and the magnetic module is only arranged at the corresponding position of the magnetic bead capturing cavity 106 by the matched instrument. The method comprises the following specific steps: and opening the third valve B3 and the sixth valve B6, and simultaneously starting the magnetic suction module by a matched instrument. Air is introduced from the sixth air channel A6, and the mixed solution in the sample processing chamber 103 is sequentially pushed into the magnetic bead capture chamber 106 and the waste liquid chamber 105. When the mixed liquid passes through the magnetic bead capture cavity 106, the magnetic beads are uniformly adsorbed on the inner wall of the magnetic bead capture cavity 106 under the action of the magnetic attraction force of the magnetic attraction module, and the rest liquid parts of the mixed liquid are all pushed into the waste liquid cavity 105, and then the third valve B3 and the sixth valve B6 are closed. The first cleaning liquid pack 123 is punctured, and the cleaning liquid is discharged from the first cleaning liquid pack 123 to the sample chamber 132 of the third flow channel A3. At this time, since the corresponding valve of the channel is in the closed state, the cleaning solution is temporarily retained in the sample solution chamber 132 and does not automatically flow into each chamber of the cartridge body 1. The fourth valve B4 and the fifth valve B5 are opened, air is introduced from the third air flow channel A3, the wash liquid is pushed into the sample processing chamber 103, and then the fourth valve B4 and the fifth valve B5 are closed. And opening a third valve B3 and a sixth valve B6, introducing air from a sixth air channel A6, and sequentially pushing the cleaning solution into the magnetic bead capturing cavity 106 and the waste liquid cavity 105 to finish the first magnetic bead cleaning. The second cleaning solution bag 125 is continuously punctured, the fifth valve B5 is opened, air is introduced from the fifth air flow channel A5, the cleaning solution is pushed into the sample processing chamber 103, and then the fifth valve B5 is closed. And opening a third valve B3 and a sixth valve B6, introducing air from a sixth air channel A6, and sequentially pushing the cleaning solution into the magnetic bead capturing cavity 106 and the waste liquid cavity 105 to finish the second magnetic bead cleaning. And continuously introducing air from the sixth air channel A6 to blow dry the magnetic bead capture cavity 106.

The second kind of magnetic bead cleaning method is to adsorb the magnetic bead and carry out a washing in sample processing chamber 103, carries out the secondary and washs in magnetic bead capture chamber 106, and supporting instrument all sets up magnetism and inhales the module in magnetic bead capture chamber 106 and sample processing chamber 103 corresponding position department. The method comprises the following specific steps: the matched instrument starts the magnetic module at the corresponding position of the sample processing cavity 103, opens the third valve B3 and the sixth valve B6, and introduces air from the sixth air passage A6. Under the magnetic attraction of the magnetic module, the magnetic beads are uniformly adsorbed on the inner wall of the sample processing chamber 103, and the rest of the liquid of the mixed liquid is completely pushed into the waste liquid chamber 105, and then the third valve B3 and the sixth valve B6 are closed. The first cleaning solution bag 123 is punctured, so that the cleaning solution flows out from the first cleaning solution bag 123 to the sample chamber 132 of the third flow channel A3. At this time, since the corresponding valve of the channel is in the closed state, the cleaning solution is temporarily retained in the sample solution chamber 132 and does not automatically flow into each chamber of the cartridge body 1. The fourth valve B4 and the fifth valve B5 are opened, and air is introduced from the third air flow channel A3 to push the wash solution into the sample processing chamber 103. Then the magnetic module is closed by the matching instrument, air is continuously introduced from the third air flow channel A3, bubbles continuously generated from the bottom of the sample processing chamber 103 are utilized to fully mix the cleaning solution and the magnetic beads, and then the fourth valve B4 and the fifth valve B5 are closed. The magnetic module located at the corresponding position of the magnetic bead capture cavity 106 is opened by the matching instrument, and the third valve B3 and the sixth valve B6 are opened to introduce air from the sixth air channel A6. Under the magnetic attraction of the magnetic module, the magnetic beads are uniformly adsorbed on the inner wall of the magnetic bead capturing cavity 106, the rest liquid part of the mixed liquid is completely pushed into the waste liquid cavity 105, and then the third valve B3 and the sixth valve B6 are closed. The second cleaning solution bag 125 is continuously punctured, the fifth valve B5 is opened, air is introduced from the fifth air flow channel A5, the cleaning solution is pushed into the sample processing chamber 103, and then the fifth valve B5 is closed. And opening a third valve B3 and a sixth valve B6, introducing air from a sixth air channel A6, and sequentially pushing the cleaning solution into the magnetic bead capturing cavity 106 and the waste liquid cavity 105 to finish the second magnetic bead cleaning. And continuously introducing air from the sixth air channel A6 to blow dry the reagent remained on the surface of the magnetic bead in the magnetic bead capturing cavity 106.

Seventh step, releasing the eluent: the eluent bag 124 is punctured, so that the eluent flows out of the eluent bag 124 to the test solution cavity 132 of the fourth gas flow channel A4. At this time, since the needle valve corresponding to the channel is in the closed state, the eluent is temporarily retained in the sample chamber 132 and does not automatically flow into each chamber of the cartridge body 1. And opening a seventh valve B7 and an eighth valve B8, introducing air from the fourth air flow channel A4, and sequentially pushing the eluent into the magnetic bead capturing cavity 106 and the mixing cavity 107. And continuously introducing air from the fourth air flow channel A4, mixing the eluent with the freeze-dried beads in the blending cavity 107 by utilizing the air bubbles continuously generated from the bottom of the blending cavity 107, and then closing the seventh valve B7 and the eighth valve B8.

Eighth step, pushing the eluent into the PCR cavity: the ninth valve B9 is opened, and air is introduced from the seventh air passage A7 to push the eluent in the mixing cavity 107 into the PCR cavity 109. Because the waterproof and breathable film 166 is arranged at the PCR cavity 109, when the four amplification cavities 162 are filled, the mixed solution stops flowing into the PCR cavity 109, and the ninth valve B9 is closed.

Ninth step, liquid flow separation: the pressurizing module of the matching instrument is used to press the first partition M1 and the second partition M2, so that the PCR cavity 109 is separated from other flow channels.

Step ten, performing an amplification cycle: the 4 amplification zones 162 of the PCR chamber 109 are subjected to temperature rise and decrease cycles by an amplification heating module of a matching instrument, and the amplification result is detected by an optical detection module.

Step ten, sealing a flow passage: after the experiment is finished, the heating module of the matched instrument is utilized to heat the wax in the wax valve cavity (blocking the cavity 108) so that the wax is melted and naturally falls down under the action of gravity, and the runner between the uniformly mixing cavity 107 and the PCR cavity 109 is blocked after solidification, so that the effect of sealing the PCR cavity 109 is achieved, and the phenomenon that an amplification product in the PCR cavity leaks out of the card box through the runner to pollute the instrument or the environment is avoided.

In summary, the present invention provides a cartridge, which reasonably arranges necessary reaction chambers such as a sample chamber, a magnetic bead storage chamber 104, a magnetic bead capture chamber 106, a waste liquid chamber 105, a mixing chamber 107 and a PCR chamber 109 on a cartridge body 1, and independently seals reagents required for reaction in a reagent pack attached to the cartridge body 1, thereby solving the problem of sealed storage of the reagents. Preferably, thermoplastic elastic glue is used as the bale breaking piece 4 to break the reagent bag.

In addition, the card box body 1 is reasonably provided with an air flow channel, a main flow channel and a valve (a specially designed thimble valve), the flow direction and the stop position of the sample and the reagent are controlled by the valve, and the flow of the liquid in the card box body 1 is controlled by the way that the air pump drives the liquid, so that the reagent bag is selectively communicated with the reaction cavity, and the control is simple and reliable.

Also, when the liquid in the cartridge is controlled using the air pump, or when the liquid reacts in the reaction chamber, bubbles are easily generated. The prior art uses cartridges of horizontal construction, where the bubbles, which are generated when pushed by a reaction or air pump, are not easily removed. The card box of the invention is of a vertical structure, and air bubbles generated in the reaction can rise from bottom to top and can be easily eliminated. And a waterproof air-permeable membrane 166 is arranged at the PCR cavity 109, so that the mixed solution of the eluent and the freeze-dried beads can remove air bubbles before entering the PCR cavity 109, and the mixed solution entering the PCR cavity 109 is ensured not to contain air bubbles.

Therefore, the card box and the detection device provided by the invention integrate the extraction, amplification and detection of the molecular diagnosis nucleic acid into a whole, and realize the automatic and closed detection process. The device is used for detection, the detection time is less than one hour, the manual operation does not exceed 2 minutes, and the device can be used for detecting pathogen genome targets of various human clinical samples and has wide applicability. The preservation and the transportation of this application device are normal atmospheric temperature condition, need not the cold chain, and the economic nature is high. The device provided by the invention has the advantages that the card box is totally closed in the detection process, the detection result is completely consistent with that of a conventional method, the operation can be carried out without the training of professional personnel, and the safety and convenience are realized.

The above description is only a preferred embodiment of the present invention, and it should be noted that, for those skilled in the art, various modifications and substitutions can be made without departing from the technical principle of the present invention, and these modifications and substitutions should also be regarded as the protection scope of the present invention.

Claims

1. A card box, which is characterized by comprising a card box body (1), a reagent pack group (2) and a sealing cover (3);

the card box body (1) is provided with a main flow channel, a valve group, a reaction cavity and an airflow channel; the valve group comprises a plurality of valves arranged at the preset positions of the main flow channel, so that one ends of the reaction cavity and the air flow channel are selectively communicated with the main flow channel, and the other end of the air flow channel is provided with an air vent for connecting an air pump;

the reaction cavity comprises a sample cavity, a magnetic bead storage cavity (104), a magnetic bead capture cavity (106), a waste liquid cavity (105), a mixing cavity (107) and a PCR cavity (109); the sample cavity is respectively communicated with the magnetic bead storage cavity (104) and the first end of the magnetic bead capture cavity (106); the first end of the magnetic bead capture cavity (106) is also communicated with the uniform mixing cavity (107), and the opposite second end of the magnetic bead capture cavity is communicated with the waste liquid cavity (105); the mixing cavity (107) is communicated with the PCR cavity (109); the sample cavity is provided with a sample adding opening (111), and the cover (3) is used for sealing the sample adding opening (111);

the reagent pack group (2) is attached to the surface of the card box body (1), and the reagent pack group (2) comprises at least four independent sealed reagent pack units;

the air flow channels at least comprise a first air flow channel (A1), a second air flow channel (A2), a third air flow channel (A3) and a fourth air flow channel (A4) which respectively correspond to the four reagent pack units; after the reagent pack unit is broken, the built-in test solution correspondingly flows into the first air flow channel (A1), the second air flow channel (A2), the third air flow channel (A3) and the fourth air flow channel (A4), and flows into one or more of the reaction chambers along the main flow channel under the control of the valve group and the air pump.

2. The cartridge of claim 1, wherein four reagent pack units are a lysis solution pack (121), a binding solution pack (122), a first wash solution pack (123), and an elution solution pack (124) corresponding to the first gas flow channel (A1), the second gas flow channel (A2), the third gas flow channel (A3), and the fourth gas flow channel (A4), respectively;

the first gas flow channel (A1) is communicated with the magnetic bead storage cavity (104), the second gas flow channel (A2) is communicated with the sample cavity, the third gas flow channel (A3) is communicated with the first end of the sample cavity or the magnetic bead capture cavity (106), and the fourth gas flow channel (A4) is communicated with the second end of the magnetic bead capture cavity (106).

3. The cartridge of claim 2, wherein the reagent pack set (2) further comprises a second wash solution pack (125), the flow channel further comprises a fifth flow channel (A5) corresponding to the second wash solution pack (125), the fifth flow channel (A5) being in communication with the first end of the sample chamber or the magnetic bead capture chamber (106).

4. A cartridge according to claim 1, wherein the cartridge body (1) is further provided with a sixth air passage (A6), a seventh air passage (A7) and an eighth air passage (A8); the one end of sixth air flue (A6), seventh air flue (A7) and eighth air flue (A8) all has the vent with the air pump is connected, the other end of sixth air flue (A6) with the top intercommunication in sample chamber, the other end of seventh air flue (A7) with mixing chamber (107) intercommunication, the other end of eighth air flue (A8) with waste liquid chamber (105) intercommunication.

5. The cartridge of claim 1, wherein the sample chamber comprises a sample addition chamber (101), a reference freeze-dried bead chamber (102), and a sample processing chamber (103), and the sample addition chamber (101) and the sample processing chamber (103) are communicated by a siphon elbow (110); one end of the internal reference freeze-dried bead cavity (102) is communicated with the bottom of the sample adding cavity (101), and the other end of the internal reference freeze-dried bead cavity is communicated with one end of the siphon elbow (111); the other end of the siphon elbow (110) is communicated with the top of the sample processing cavity (103); the sample adding port (111) is arranged at the top of the sample adding cavity (101).

6. The cartridge of claim 1, wherein the reaction chamber further comprises a blocking chamber (108); the blocking cavity (108) is connected to the main flow channel between the blending cavity (107) and the PCR cavity (109) so as to block the communication between the blending cavity (107) and the PCR cavity (109) when a preset condition is met.

7. Cartridge according to claim 1, wherein the cartridge body (1) comprises a cartridge plate (11), a front membrane (12) and a rear membrane (13); the front surface of the cassette board (11) is provided with a concave cavity or a through hole, the front film (12) is attached to the front surface of the cassette board (11), and the rear film (13) is attached to the back surface of the cassette board (11) to seal the concave cavity or the through hole, so that the reaction cavity is formed.

8. The cartridge of claim 7, wherein the primary flow channel is comprised of at least one positive and negative channel structure; the front groove structure and the back groove structure comprise a front groove (N1), a back groove (N2) and a communication hole (N3); the front groove (N1) is arranged on the front surface of the cassette plate (11), the back groove (N2) is arranged on the back surface of the cassette plate (11), and the communication hole (N3) is used for communicating the front groove (N1) with the back groove (N2); the front film (12) and the rear film (13) seal the front groove (N1), the back groove (N2), and the communication hole (N3) to form the main flow passage.

9. The cartridge of claim 8, wherein the PCR chamber (109) comprises a central chamber (161), an amplification chamber (162), a zone flow channel (163), a central flow channel groove (164), a gas containing zone (165), a water-proof gas-permeable membrane (166), and a zone gas channel (167);

the central flow channel groove (164) is arranged on the front surface of the central cavity (161), the main flow channel is arranged on the back surface of the central cavity (161), the main flow channel is communicated with one end of the central flow channel groove (164), and the other end of the central flow channel groove (164) is communicated with the amplification cavity (162) through the zone flow channel (163); the waterproof breathable film (166) is attached to the central cavity (161) to seal the central runner groove (164); one end of the area air channel (167) is arranged at the edge of the sealing area of the waterproof breathable film (166), and the other end of the area air channel is communicated with the air containing area (165).

10. The cartridge of claim 8, wherein the valve comprises a valve cavity (151), an elastic pad (152) and at least one protrusion (153);

the side surface of the valve cavity (151) is provided with a first flow passage hole (154), and the bottom of the valve cavity is provided with a second flow passage hole (155); the elastic pad (152) is located above the second flow passage hole (155); the convex part (153) is arranged on the surface of the elastic cushion (152) facing the valve cavity (151) and/or is arranged on the surface of the valve cavity (151) at a position corresponding to the elastic cushion (152) so that a gap is formed between the elastic cushion (152) and the second flow passage hole (155).

11. The cartridge of claim 10, wherein the second flow passage hole (155) is provided at the center of the valve cavity (151); one convex part (153) is arranged, and the convex part (153) is arranged on the surface of the valve cavity (151) and is positioned on the connecting line of the first flow passage hole (154) and the second flow passage hole (155).

12. The cartridge of claim 11, wherein the first flow channel hole (154) is provided at an upper portion of a side of the valve chamber (151) and a corresponding lower portion is inclined downward from the outside to the inside.

13. A cartridge according to claim 1, wherein the set of reagent packs comprises a housing (21), a sealing membrane (22) and an attachment (23);

the shell (21) is provided with at least four liquid containing cavities with openings facing the front side, corresponding test solutions are preset in the liquid containing cavities, and the sealing film (22) is attached to the front side of the shell (21) to seal the liquid containing cavities, so that corresponding reagent pack units are formed; one surface of the attachment (23) is used for being connected with the front surface of the shell (21), the other surface of the attachment is used for being connected with the surface of the cassette board (11), and a hollow part is formed at the position, corresponding to the liquid containing cavity, on the attachment (23).

14. The cartridge of claim 13, wherein the first air flow channel (A1), the second air flow channel (A2), the third air flow channel (A3), and the fourth air flow channel (A4) each include an upper air passage (131), a test solution chamber (132), and a lower air passage (133); one end of the upper air channel (131) is provided with the vent, and the other end of the upper air channel is communicated with the upper part of the test solution cavity (132); one end of the lower flow channel (133) is communicated with the lower part of the test solution cavity (132), and the other end of the lower flow channel is connected into the main flow channel; the test solution cavity (132) corresponds to the solution containing cavity, and the built-in test solution flows into the test solution cavity (132) and the lower runner (133) in sequence after the reagent pack unit is broken.

15. A cartridge according to claim 14, further comprising a bursting member (4); the bale breaking piece (4) is thermoplastic elastic glue;

the edge of test solution chamber (132) is equipped with groove (134) of moulding plastics, the middle part department of test solution chamber (132) is equipped with parting strip (135) will test solution chamber (132) is separated into the epicoele and the lower chamber of intercommunication each other, be equipped with on the parting strip (135) on the surface with mould plastics the separation of groove (134) intercommunication and mould plastics groove (136), and all test solution chamber (132) mould plastics groove (134) and separate and mould plastics groove (136) intercommunication each other and form a runner of moulding plastics, and thermoplastic elastic material is in mould plastics and become in the runner thermoplastic elastic glue.

16. A cartridge according to claim 1, wherein the cover (3) comprises a dosing plate (31) and a plug (32);

the sample adding plate (31) is provided with a sample adding opening (111) and a first fixing part (311); the sample adding plate (31) is arranged at the top of the sample cavity;

the plug cap (32) comprises a base plate (321), a plunger (322) and a second fixing part (323); one end of the plunger (322) is arranged on the bottom surface of the base plate (321), and the other end of the plunger is inserted into the sample adding opening (111) to seal the sample adding opening (111);

one end of the second fixing part (323) is connected with the side surface of the substrate (321) through a bendable piece (324), and the other end of the second fixing part is connected with the first fixing part (311).

17. A testing device comprising a cartridge according to any one of claims 1 to 16 and associated apparatus.