A cartridge and detection device

Abstract

The application relates to the field of biological medical technology and discloses a card box and a detection device, wherein the card box comprises a card box body, a reagent bag group, a cover and a waterproof and breathable film; a main flow channel, a valve group, a reaction cavity and an airflow channel are arranged on the card box body; the reagent bag group is attached to the surface of the card box body, and the reagent bag group comprises at least four independently sealed reagent bag units; after the reagent bag units are broken, the built-in test solution flows into the first airflow channel, the second airflow channel, the third airflow channel and the fourth airflow channel, 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 application can realize automatic processing of nucleic acid extraction, amplification and detection, only needs a small number of manual operation steps, is simple to operate, safe and convenient, has high sealing performance, and is more accurate in detection results.

Description

Technical Field

[0001] This invention relates to the field of biomedical technology, and in particular to a card holder and a detection device. Background Technology

[0002] Nucleic acid testing plays a vital role in many fields of biochemical analysis and has been widely applied in the biomedical field.

[0003] Current technologies typically use centrifugation column methods or magnetic bead methods to extract nucleic acids from samples, generally requiring four steps: lysis, binding, washing, and elution. After nucleic acid extraction, subsequent steps such as nucleic acid hybridization, polymerase chain reaction (PCR), and bioarrays are performed to complete nucleic acid detection.

[0004] Because nucleic acid testing involves many steps, it is extremely difficult to achieve fully automated testing equipment "from sample to result". Furthermore, in terms of the transfer of effective components in each step, existing technologies mostly use manual transfer methods, which are not only cumbersome, time-consuming and labor-intensive, but also difficult to transfer materials fully and efficiently. Manual operation is prone to causing unstable results, making the testing difficult to implement.

[0005] In addition, the mainstream technology for molecular detection is quantitative real-time PCR. Due to the exponential amplification of templates by PCR technology, the existing open consumables make the entire operation process prone to PCR aerosol contamination, affecting the purity of the extract, thus limiting the further clinical application of quantitative real-time PCR technology.

[0006] Therefore, existing technologies urgently need improvement. Summary of the Invention

[0007] The purpose of this invention is to provide a cartridge and a detection device to solve the technical problems of existing nucleic acid detection technologies, such as the difficulty in achieving fully automated processing, and the instability, low accuracy, and low efficiency of detection results caused by manual operation and the use of open consumables.

[0008] To achieve the above objectives, the present invention provides a card box, comprising a card box body, a reagent pack assembly, a cap, and a waterproof and breathable membrane;

[0009] The cartridge body is provided with a main channel, a valve group, a reaction chamber and an airflow channel; the valve group includes a number of valves located at predetermined positions in the main channel, so that one end of the reaction chamber and one end of the airflow channel can be selectively connected to the main channel, and the other end of the airflow channel has a vent for connecting an air pump, and the waterproof and breathable membrane is used to seal the vent.

[0010] The reaction chamber includes a sample chamber, a magnetic bead storage chamber, a magnetic bead capture chamber, a waste liquid chamber, a mixing chamber, a first quantitative chamber, and a PCR chamber. The sample chamber is connected to the first ends of both the magnetic bead storage chamber and the magnetic bead capture chamber. The first end of the magnetic bead capture chamber is also connected to the mixing chamber, and its opposite second end is connected to the waste liquid chamber. The mixing chamber is connected to the first end of the first quantitative chamber, and its opposite second end is connected to the PCR chamber. The sample chamber has a sample dispensing port, and the cap is used to seal the sample dispensing port.

[0011] The reagent pack assembly is affixed to the surface of the cartridge body, and the reagent pack assembly includes at least four independently sealed reagent pack units; the four reagent pack units are respectively a lysis buffer pack, a binding buffer pack, a first washing buffer pack, and an elution buffer pack;

[0012] The airflow channels include at least a first airflow channel, a second airflow channel, a third airflow channel, and a fourth airflow channel, which correspond to the four reagent pack units respectively. After the reagent pack unit is broken, the test liquid inside flows into the first airflow channel, the second airflow channel, the third airflow channel, and the fourth airflow channel respectively, 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.

[0013] In some embodiments of this application, the first gas flow channel corresponds to the lysis buffer package, and the second gas flow channel corresponds to the binding buffer package; the main flow channel includes a first branch channel and a second branch channel;

[0014] The two ends of the first branch channel are respectively connected to the bottom end of the sample cavity and the bottom end of the first airflow channel;

[0015] One end of the second branch channel is connected to the bottom of the sample cavity, and the other end is connected to the first branch channel. This connection point is the first connection point. Both ends of the magnetic bead storage cavity are respectively connected to the second branch channel.

[0016] The bottom end of the second airflow channel is connected to the pipe section on the first branch channel located between the first airflow channel and the first connection point;

[0017] The valves are provided at predetermined positions in the first branch channel and the second branch channel to control the lysis solution in the lysis solution package and the binding solution in the binding solution package to flow directly into the sample cavity or flow sequentially through the magnetic bead storage cavity and the sample cavity.

[0018] In some embodiments of this application, the reagent package group further includes a second cleaning solution package and a reserve solution package; the airflow channel further includes a fifth airflow channel and a sixth airflow channel;

[0019] The third airflow channel corresponds to the first cleaning fluid pack, and the bottom end of the third airflow channel is connected to the pipe section on the first branch channel located between the second airflow channel and the first connection point;

[0020] The fifth airflow channel corresponds to the second cleaning fluid pack, and the bottom end of the fifth airflow channel is connected to the pipe section on the first branch channel located between the third airflow channel and the first connection point;

[0021] The sixth airflow channel corresponds to the reserved liquid package, and the bottom end of the sixth airflow channel is connected to the pipe section on the first branch channel located between the fifth airflow channel and the first connection point.

[0022] In some embodiments of this application, the reaction chamber further includes a second metering chamber; the top of the second metering chamber is provided with a seventh gas channel, and the other end of the seventh gas channel has the vent; the bottom of the second metering chamber is connected to a pipe section on the first branch channel located between the sixth gas channel and the first connection point, and the valves are respectively provided on both sides of the connection point.

[0023] In some embodiments of this application, the main channel further includes a third branch channel, a predetermined section of which forms the magnetic bead capture cavity; the first end of the third branch channel is connected to the first connection point, and its opposite second end is connected to one end of the waste liquid cavity, and the other end of the waste liquid cavity is provided with an outlet channel;

[0024] The fourth airflow channel corresponds to the eluent pack, and the bottom end of the fourth airflow channel is connected to the second end of the third branch channel;

[0025] The valve is provided at a predetermined position in the third branch channel to control the liquid in the magnetic bead capture chamber to flow from the second end of the third branch channel into the waste liquid chamber, and to control the eluent in the eluent package to flow from the second end of the third branch channel into the magnetic bead capture chamber.

[0026] In some embodiments of this application, the sample cavity includes a sample inlet cavity and a sample processing cavity, and the sample inlet cavity and the sample processing cavity are connected by a siphon bend.

[0027] One end of the siphon bend is connected to the bottom of the sample inlet chamber, and the other end is connected to the top of the sample processing chamber; the top of the sample inlet chamber is provided with the sample dispensing port; the top of the sample processing chamber is provided with an eighth air passage.

[0028] In some embodiments of this application, a condensation chamber is provided at a predetermined position of the eighth airway, and a desiccant is pre-placed in the condensation chamber.

[0029] In some embodiments of this application, the main channel further includes a fourth branch channel, a fifth branch channel, and a sixth branch channel;

[0030] One end of the fourth branch channel is connected to the bottom of the sample processing chamber, and the other end is connected to the top of the mixing chamber.

[0031] One end of the fifth branch channel is connected to the first end of the third branch channel, and the other end is connected to the first end of the mixing chamber;

[0032] One end of the sixth branch channel is connected to the opposite second end of the mixing chamber, and the other end is connected to the PCR chamber; the top of the PCR chamber is provided with a tenth airway;

[0033] The top of the first quantitative chamber is provided with a ninth airway, and the other end of the ninth airway is provided with the air inlet; the bottom of the first quantitative chamber is connected to the tube segment located between the mixing chamber and the PCR chamber on the sixth branch channel, and the valves are respectively provided on both sides of the connection point.

[0034] In some embodiments of this application, several PCR chambers are arranged side by side, and each PCR chamber has a valve at its bottom that allows it to selectively communicate with the first quantitative chamber.

[0035] In some embodiments of this application, the valve is a needle valve; the needle valve has an internal channel, and a sealing membrane is provided on the channel. When the sealing membrane is subjected to a predetermined pressure, it deforms, thereby sealing the channel.

[0036] In some embodiments of this application, the reagent kit assembly includes a shell and a sealing film; the shell is provided with at least four liquid-containing cavities, each containing a corresponding test solution, and a groove is provided around the edge of each liquid-containing cavity; the number of sealing films corresponds to the number of liquid-containing cavities, and the sealing films are affixed to the grooves to seal the liquid-containing cavities, thereby forming the reagent kit unit.

[0037] In some embodiments of this application, the airflow channel includes an upper airflow channel, a test liquid chamber, and a lower flow channel; one end of the upper airflow channel has the vent, and the other end is connected to the upper part of the test liquid chamber; one end of the lower flow channel is connected to the lower part of the test liquid chamber, and the other end is connected to the main flow channel; the test liquid chamber corresponds to the liquid-containing chamber, and after the reagent pack unit is broken, the test liquid inside it flows into the test liquid chamber and the lower flow channel in sequence.

[0038] In some embodiments of this application, a puncture needle is provided at a predetermined distance between the test solution chamber and the sealing film.

[0039] In some embodiments of this application, the card holder body includes a card holder plate, a front film, and a rear film;

[0040] The front side of the cartridge plate has several recesses, and the front membrane is attached to the front side of the cartridge plate to seal the recesses, thereby forming the reaction chambers other than the PCR chamber.

[0041] The card tray plate has several through holes. The front membrane is attached to the front side of the card tray plate, and the rear membrane is attached to the back side of the card tray plate to seal the through holes, thereby forming the PCR chamber.

[0042] The card holder plate (11) is provided with a plurality of grooves, and the front film is attached to the front side of the card holder plate to seal the grooves, thereby forming the airflow channel and the main flow channel.

[0043] This application also proposes a testing device, including a card box and supporting instruments as described in any of the preceding claims.

[0044] Compared with the prior art, the advantages of the card holder and detection device of this invention are as follows:

[0045] The cartridge and detection device of this invention arranges each reaction chamber on the cartridge body and independently seals the reagents required for the reaction in a reagent pack attached to the cartridge body. The supporting instrument controls the main flow channel, air flow channel and valve on the cartridge body, so that the reagent pack and the reaction chamber are selectively connected, thereby realizing the automated processing of nucleic acid extraction, amplification and detection. Only a few manual operation steps are required, the operation is simple, safe and convenient, the sealing is strong, and the detection results are more accurate. Attached Figure Description

[0046] To more clearly illustrate the technical solutions in the embodiments of the present invention, the drawings used in the description of the embodiments or the prior art will be briefly introduced below. Obviously, the drawings described below are only some embodiments of the present invention. For those skilled in the art, other drawings can be obtained based on these drawings without creative effort.

[0047] Figure 1 This is an exploded view of the card box according to an embodiment of the present invention;

[0048] Figure 2 This is a front structural diagram of the card box and the central rod of the supporting instrument;

[0049] Figure 3 This is a schematic diagram of the front structure of the card holder with the front film removed;

[0050] Figure 4 This is a schematic diagram of the back structure of the card box;

[0051] Figure 5 This is a side view of the card box.

[0052] Figure 6 This is a top view of the card box.

[0053] Figure 7 This is a schematic diagram of the axonometric structure of the card box plate;

[0054] Figure 8 This is a schematic diagram of the front structure of the card box panel;

[0055] Figure 9 This is a schematic diagram of the back structure of the card box panel;

[0056] Figure 10 yes Figure 2 D-D sectional view;

[0057] Figure 11 This is a schematic diagram of the engagement state between the center rod and the ejector valve. Figure 1 ;

[0058] Figure 12 This is a schematic diagram of the engagement state between the center rod and the ejector valve. Figure 2 ;

[0059] Figure 13 This is a schematic diagram of the axial structure of the housing;

[0060] Figure 14 This is a front view structural diagram of the shell;

[0061] Figure 15 This is a side view of the shell structure.

[0062] Figure 16 This is a top view of the shell structure.

[0063] Figure 17 yes Figure 3 Middle E-E sectional view;

[0064] Figure 18 yes Figure 17 Enlarged image in E1;

[0065] Figure 19 This is a schematic diagram of the structure of the mixing chamber, the first quantitative chamber, and the PCR chamber;

[0066] In the diagram, 1. Cartridge body; 11. Cartridge plate; 12. Front membrane; 13. Rear membrane; 2. Reagent pack; 21. Shell; 22. Sealing film; 23. Liquid chamber; 24. Adhesive groove; 3. Cap; 4. Waterproof and breathable membrane; 5. Sample dispensing port; 6. Pulse valve; 61. Sealing film; 7. Positioning pin; 8. Center rod; 9. Puncture needle; 91. Needle tip;

[0067] 101. Sample addition chamber; 102. Siphon bend; 103. Sample processing chamber; 104. Cooling chamber; 105. Magnetic bead storage chamber; 106. Second quantitative chamber; 107. Waste liquid chamber; 108. Magnetic bead capture chamber; 109. Mixing chamber; 110. First quantitative chamber; 111. PCR chamber 1; 112. PCR chamber 2; 113. PCR chamber 3; 114. PCR chamber 4;

[0068] 121. Lysis buffer pack; 122. Binding buffer pack; 123. First washing buffer pack; 124. Elution buffer pack; 125. Second washing buffer pack; 126. Reserve buffer pack;

[0069] A1, First airflow channel; A2, Second airflow channel; A3, Third airflow channel; A4, Fourth airflow channel; A5, Fifth airflow channel; A6, Sixth airflow channel; A7, Seventh airflow channel; A8, Eighth airflow channel; A9, Ninth airflow channel; A10, Tenth airflow channel;

[0070] 131. Upper airway; 132. Test liquid chamber; 133. Lower flow channel;

[0071] 141. First branch channel; 142. Second branch channel; 143. Third branch channel; 144. Fourth branch channel; 145. Fifth branch channel; 146. Sixth branch channel;

[0072] 1501, Gas Channel; 1502, Liquid Channel; 151, Channel 1; 152, Channel 2; 153, Channel 3; 154, Channel 4. Detailed Implementation

[0073] The specific embodiments of the present invention will be described in further detail below with reference to the accompanying drawings and examples. The following examples are for illustrative purposes only and are not intended to limit the scope of the invention.

[0074] In the description of this invention, it should be understood that the terms "upper", "lower", "left", "right", "top", "bottom", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing this invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on this invention.

[0075] It should be understood that the terms "length", "width", "upper", "lower", "front", "rear", "left", "right", "vertical", "horizontal", "top", "bottom", "inner", "outer", etc., indicate the orientation or positional relationship based on the orientation or positional relationship shown in the accompanying drawings. They are only for the convenience of describing the present invention and simplifying the description, and do not indicate or imply that the device or element referred to must have a specific orientation, or be constructed and operated in a specific orientation. Therefore, they should not be construed as limitations on the present invention.

[0076] Furthermore, the terms "first" and "second" are used for descriptive purposes only and should not be construed as indicating or implying relative importance or implicitly specifying 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 this invention, "a plurality of" means two or more, unless otherwise explicitly specified.

[0077] See Figure 1 -6 is a card box according to a preferred embodiment of the present invention, including a card box body 1, a reagent pack 2, a cap 3 (rubber stopper cap), a waterproof and breathable membrane 4, and a positioning pin 7.

[0078] The cartridge body 1 is provided with a main channel, a valve assembly, a reaction chamber, and an airflow channel. The valve assembly includes several valves located at predetermined positions on the main channel, allowing one end of the reaction chamber and the airflow channel to selectively connect to the main channel. The other end of the airflow channel has a vent for connecting an air pump. A waterproof and breathable membrane 4 is used to seal the vent using hot-press welding technology.

[0079] The reaction chamber includes at least a sample chamber, a magnetic bead storage chamber 105, a magnetic bead capture chamber 108, a waste liquid chamber 107, a mixing chamber 109, a first quantitative chamber 110, and a PCR chamber. The sample chamber is connected to the first ends of both the magnetic bead storage chamber 105 and the magnetic bead capture chamber 108. The first end of the magnetic bead capture chamber 108 is also connected to the mixing chamber 109, and its opposite second end is connected to the waste liquid chamber 107. The mixing chamber 109 is connected to the first end of the first quantitative chamber 110, and the opposite second end of the first quantitative chamber 110 is connected to the PCR chamber. The sample chamber has a sample dispensing port 5, and a cap 3 is used to seal the sample dispensing port 5.

[0080] The reagent pack assembly 2 is affixed to the surface of the cartridge body 1, and the reagent pack assembly 2 includes at least four independently sealed reagent pack units. The four reagent pack units are a lysis buffer pack 121, a binding buffer pack 122, a first washing buffer pack 123, and an elution buffer pack 124.

[0081] The airflow channels include at least a first airflow channel A1, a second airflow channel A2, a third airflow channel A3, and a fourth airflow channel A4, which correspond to the four reagent pack units respectively. After the reagent pack unit is broken, the test solution inside flows into the first airflow channel A1, the second airflow channel A2, the third airflow channel A3, and the fourth airflow channel A4 respectively, and flows into one or more of the reaction chambers along the main flow channels under the control of the valve group and the air pump.

[0082] Based on the above description, this application proposes a preferred embodiment of a card holder as follows:

[0083] See Figure 1 -6. The cartridge body 1 includes a cartridge plate 11, a front film 12, and a rear film 13. The front side of the cartridge plate 11 has recesses, through holes, or grooves. The front film 12 is attached to the front side of the cartridge plate 11, and the rear film 13 is attached to the back side of the cartridge plate 11, sealing the recesses, through holes, or grooves to form various reaction chambers, main flow channels, and airflow channels. Specifically, the front film 12 and the rear film 13 can be made of polypropylene plastic film and are attached to the surface of the cartridge plate 11 by laser welding. In the following description, all descriptions of left, right, top, and bottom orientations are based on the front side of the cartridge plate 11.

[0084] See Figure 7 -9. The upper right of the cartridge plate 11 is equipped with a sample addition chamber 101, a siphon bend 102, a sample processing chamber 103, a condensation chamber 104, a magnetic bead storage chamber 105, and a second quantitative chamber 106. The lower right is equipped with a mixing chamber 109, a first quantitative chamber 110, and four PCR chambers: PCR chamber 111 (No. 1), PCR chamber 112 (No. 2), PCR chamber 113 (No. 3), and PCR chamber 114 (No. 4). The upper left is equipped with a reagent pack group 2. The lower left is equipped with a waste liquid chamber 107 and a magnetic bead capture chamber 108. The magnetic bead storage chamber 105 is pre-filled with magnetic particles, such as lyophilized magnetic beads. Passing the sample through the magnetic bead storage chamber 105 is primarily for extracting biomolecules such as nucleic acids using magnetic particles, including magnetic beads; this is a common method for nucleic acid extraction in existing technologies. The mixing chamber 109 is pre-filled with lyophilized beads for mixing with the sample and elution buffer; lyophilized beads commonly used in existing nucleic acid detection technologies can be used. The sample processing chamber 103 is pre-filled with a drying reagent. After thorough mixing of the drying reagent and the sample, it aids in the digestion of potential nucleases in the sample. The drying reagent in this application can be any drying reagent conventionally used in existing nucleic acid detection technologies that can achieve the above-mentioned function.

[0085] The sample inlet chamber 101 is located at the upper right of the cartridge plate 11. One end of the siphon bend 102 connects to the bottom of the sample inlet chamber 101, and the other end connects to the top of the sample processing chamber 103. The siphon bend 102 has two downward bends and two upward bends. Furthermore, the sample processing chamber 103 contains a desiccant. An eighth airway A8 is located at the top of the sample processing chamber 103, and a condensation chamber 104 is located at a predetermined position in the eighth airway A8. The condensation chamber 104 is located on the left side of the sample processing chamber 103 and contains a desiccant pre-placed within it to absorb moisture from the sample.

[0086] Those skilled in the art should understand that the specific locations of the reaction chambers described above are only preferred embodiments and not the only options.

[0087] See Figure 3 -6. Reagent pack group 2 is attached to the upper left back of the cartridge plate 11. Reagent pack group 2 includes six independently sealed reagent pack units: eluent pack 124, lysis buffer pack 121, binding buffer pack 122, first cleaning buffer pack 123, second cleaning buffer pack 125, and reserve buffer pack 126 (the labels in the figure only represent the location of the reagent pack units and do not represent the complete structure of the reagent pack units). From left to right, the upper left of the cartridge plate 11 is provided with a fourth airflow channel A4, a first airflow channel A1, a second airflow channel A2, a third airflow channel A3, a fifth airflow channel A5, and a sixth airflow channel A6, corresponding to the eluent pack 124, lysis buffer pack 121, binding buffer pack 122, first cleaning buffer pack 123, second cleaning buffer pack 125, and reserve buffer pack 126, respectively. One end of each of the above airflow channels is a gas pump port for ventilation, which is connected to an external gas pump. In the accompanying drawings of this application, for ease of understanding and labeling, the annotation lines are placed at the air pump holes of the airways. For example, the annotation line A1 in the figure is placed at the air pump hole of the first airway A1, and the same applies to the other airways.

[0088] Those skilled in the art should understand that there is a one-to-one correspondence between the reagent package units and the airflow channels, but the above-described arrangement of the reagent package units is only a preferred embodiment and not the only option.

[0089] See Figure 8 The front of the card box plate 11 is slotted and sealed by the front membrane 12 to form the main channel, which is composed of several branch channels.

[0090] The two ends of the first branch channel 141 are respectively connected to the bottom end of the sample processing chamber 103 and the bottom end of the first airflow channel A1, and the bottom end of the first airflow channel A1 is provided with a valve B11.

[0091] One end of the second branch channel 142 is connected to the bottom of the sample processing cavity 103, and the other end is connected to the first branch channel 141. This connection point is the first connection point C1. Both ends of the magnetic bead storage cavity 105 are connected to the second branch channel 142, and the pipe section between the magnetic bead storage cavity 105 and the sample processing cavity 103 is equipped with a valve B1.

[0092] The bottom end of the second airflow channel A2 is connected to the pipe section located between the first airflow channel A1 and the first connection point C1 on the first branch channel 141 through valve B10.

[0093] The bottom end of the third airflow channel A3 is connected to the pipe section located between the second airflow channel A2 and the first connection point C1 on the first branch channel 141 via valve B9.

[0094] The bottom end of the fifth airflow channel A5 is connected to the pipe section located between the third airflow channel A3 and the first connection point C1 on the first branch channel 141 via valve B8.

[0095] The bottom end of the sixth airflow channel A6 is connected to the pipe section located between the fifth airflow channel A5 and the first connection point C1 on the first branch channel 141 via a pin valve.

[0096] The top of the second metering chamber 106 is provided with a seventh air passage A7, and the other end of the seventh air passage A7 has an air inlet for connecting to an air pump. The bottom of the second metering chamber 106 is connected to a pipe section on the first branch passage 141 located between the sixth air passage A6 and the first connection point C1, and a valve B6 is provided on the left side and a valve B5 is provided on the right side of this connection point.

[0097] The predetermined section of the third branch channel 143 forms the magnetic bead capture chamber 108. The first end of the third branch channel 143 is connected to the first connection point C1, and its opposite second end is connected to one end of the waste liquid chamber 107, and a first valve B12 is provided at the connection position. The other end of the waste liquid chamber 107 is provided with an air outlet, and the other end of the air outlet is connected to the fourth airflow channel A4.

[0098] The bottom end of the fourth airflow channel A4 is connected to the second end of the third branch channel 143 through the valve B13.

[0099] One end of the fourth branch channel 144 is connected to the top of the mixing chamber 109, and the other end is connected to the first branch channel 141. This connection point is the second access point C2. A valve B2 is installed between the bottom of the sample processing chamber 103 and the second access point C2. A valve B3 is installed between the first access point C1 and the second access point C2. A valve B4 is installed between the second access point C2 and the mixing chamber 109, so that the other end of the fourth branch channel 144 can be connected to the bottom of the sample processing chamber 103 through the pipe section of the first branch channel 141.

[0100] One end of the fifth branch channel 145 is connected to the first end of the third branch channel 143, and the other end is connected to the first end of the mixing chamber 109. A valve B14 is provided on the fifth branch channel 145.

[0101] One end of the sixth branch channel 146 is connected to the second end of the mixing chamber 109, and the other end is connected to the PCR chamber.

[0102] Specifically, the top of the first quantitative chamber 110 is provided with a ninth airway A9, and the other end of the ninth airway A9 has a vent. The top of the PCR chamber is provided with a tenth airway A10, and the other end of the tenth airway A10 has a vent.

[0103] See Figure 19 In this embodiment, there are four PCR chambers: PCR chamber 111, PCR chamber 212, PCR chamber 313, and PCR chamber 414. Each PCR chamber corresponds to a channel: PCR chamber 111 corresponds to channel 151, PCR chamber 212 corresponds to channel 2152, PCR chamber 313 corresponds to channel 3153, and PCR chamber 414 corresponds to channel 4154. Specifically, taking channel 4154 as an example, each of the above channels includes a gas channel 1501 and a liquid channel 1502. One end of the gas channel 1501 is connected to the top of PCR chamber 414, and the other end is connected to the tenth gas channel A10. One end of the liquid channel 1502 is connected to the bottom of PCR chamber 414, and the other end is used to connect to the sixth branch channel 146.

[0104] In this embodiment, the other end of the sixth branch channel 146 is connected to channel 4 154. The first metering chamber 110, channel 151, channel 2 152, and channel 3 153 can all be considered as connected to the sixth branch channel 146. A valve B15 is provided on the sixth branch channel 146 at the position between the mixing chamber 109 and the first metering chamber 110. Valves B16, B17, B18, and B19 are respectively provided on the liquid channels 1502 of channels 151, 252, 353, and 454. The gas channels 1501 of channels 151, 252, 353, and 454 are all connected to the tenth gas channel A10.

[0105] In the above embodiment, the cartridge plate 11 has four circular through holes of the same size. A front film 12 and a rear film 13 are respectively attached to the front and back of the cartridge plate 11 to seal the through holes, thereby forming PCR chamber 111, PCR chamber 112, PCR chamber 113, and PCR chamber 114. As shown in the figure, the four PCR chambers are arranged side-by-side, with adjacent PCR chambers staggered vertically, so that the angle between the line connecting the centers of the four PCR chambers and the horizontal line is 30°–60°, preferably 40°–50°, forming a parallelogram. This design effectively saves space for PCR chamber arrangement.

[0106] In the above embodiments, the volume of the first quantitative chamber 110 is related to the volume of the PCR chamber, and the volume of the mixing chamber 109 is also related to the total volume of all PCR chambers. Since there will be some liquid loss when passing through the flow channels and passages, it is preferable that the volume of the first quantitative chamber 110 is slightly larger than the volume of the PCR chamber, so that sufficient elution mixture can be measured at once to fill the PCR chamber. Preferably, the ratio of the volume of the first quantitative chamber 110 to the volume of the PCR chamber is between 1.1 and 1.5, so as to meet the above-mentioned measurement requirements while reducing the volume ratio of the first quantitative chamber 110 on the cartridge plate 11. Furthermore, the volume of the mixing chamber 109 is preferably set to be slightly larger than the total volume of all PCR chambers, so as to store sufficient elution mixture at once to fill all PCR chambers. Preferably, the ratio of the volume of the mixing chamber 109 to the total volume of all PCR chambers is between 1.1 and 1.5, so as to meet the above-mentioned storage requirements while reducing the volume ratio of the mixing chamber 109 on the cartridge plate 11.

[0107] In the above embodiments, all valves B1-B19 are pin valves 6. See also Figure 10 The accompanying instrument is equipped with a drive module (center rod) 8 that works in conjunction with the ejector valve 6. See 11-12. The ejector valve 6 has an internal channel with a sealing membrane 61. In its initial state, the drive module 8 and the ejector valve 6 are at a certain distance, the sealing membrane 61 is not under pressure and does not deform, the channel is unobstructed, meaning the ejector valve 6 is in the open state. Figure 11 As shown. When the drive module 8 moves forward to contact the ejector valve 6, the sealing diaphragm 61 is subjected to pressure and deforms, thereby closing the channel, that is, the ejector valve 6 is in the closed state, as shown. Figure 12 As shown.

[0108] In the above embodiments, see Figure 1 The reagent kit assembly 2 includes a shell 21 and a sealing film 22. The reagent kit assembly 2 is located on the upper left side of the cartridge plate 11. Specifically, the shell 21 is laser-welded to the surface of the cartridge plate 11. See also... Figure 13-16. The housing 21 has six liquid-containing chambers 23, each pre-filled with a corresponding test solution, and each chamber 23 has a groove 24 along its edge. The number of sealing films 22 is the same as the number of liquid-containing chambers 23, and they are correspondingly affixed to the grooves 24 of each liquid-containing chamber 23 to seal each chamber 23, thus forming independent reagent package units. The sealing films 22 can be aluminum-plastic films, and each liquid-containing chamber 23 is sealed using hot-press welding technology.

[0109] The arrangement of the liquid-containing chambers 23 on the shell 21 is as follows: there are three rows of liquid-containing chambers 23 from top to bottom. The liquid-containing chambers 23 in each row are arranged at equal intervals, and each liquid-containing chamber 23 in the even-numbered rows corresponds to the gap between each pair of adjacent liquid-containing chambers 23 in the odd-numbered rows. This arrangement of the liquid-containing chambers 23 facilitates the provision of space for air passages for each liquid-containing chamber 23, making the airflow arrangement more orderly.

[0110] Based on the above arrangement pattern, when reagent package group 2 has a total of six reagent package units, see [reference needed]. Figure 14 ( Figure 14 The positions of the liquid-containing chambers 23 are marked to indicate the reagent pack units, including the lysis buffer pack 121, the first cleaning buffer pack 123 and the reserve buffer pack 126 arranged in sequence in the first row; the binding buffer pack 122 located between the lysis buffer pack 121 and the first cleaning buffer pack 123 in the second row, and the second cleaning buffer pack 125 located between the first cleaning buffer pack 123 and the reserve buffer pack 126; and the elution buffer pack 124 corresponding to the position of the lysis buffer pack 121 in the third row.

[0111] The liquid-containing cavity 23 is rhomboid in shape, with its two ends forming upward and downward protruding pointed arcs respectively. The depth of each liquid-containing cavity 23 can be set according to the preset test liquid volume. The depth of the liquid-containing cavities 23 located in the first row and the first column is greater than that of the liquid-containing cavities 23 in other positions. In this embodiment, the depth of the lysis buffer package 121 is the largest.

[0112] Existing technologies typically perform only one cleaning cycle. However, this application, due to the structure of reagent pack group 2, makes it easier to add reagent pack units. Therefore, a second cleaning solution pack 125 is provided for secondary cleaning, resulting in better cleaning of the magnetic beads. Furthermore, reagent pack group 2 also includes a reserved solution pack 126, reserving space for future technological development or process improvements.

[0113] In the above embodiments, see Figure 17 and Figure 18Taking the sixth airflow channel A6 as an example, the airflow channel includes an upper airflow channel 131, a test liquid chamber 132, and a lower flow channel 133. One end of the upper airflow channel 131 has a vent, and the other end connects to the upper part of the test liquid chamber 132. One end of the lower flow channel 133 connects to the lower part of the test liquid chamber 132, and the other end connects to the main flow channel. The test liquid chamber 132 corresponds to the liquid-containing chamber 23. After the reagent package unit is broken, the test liquid inside flows sequentially into the test liquid chamber 132 and the lower flow channel 133, and then into the main flow channel.

[0114] In the above embodiment, a piercing needle 9 is provided at a predetermined distance from the sealing membrane 22 within the test solution chamber 132. Upon receiving a predetermined pressing pressure, the piercing needle 9 pierces the sealing membrane 22, allowing the test solution within the reagent pack unit to flow into the test solution chamber 132. Furthermore, the piercing needle 9 is an elastically deformable component, with pointed needle portions 91 at both ends facing the sealing membrane 22. The pressing module of the accompanying instrument simultaneously squeezes the pointed needle portions 91 at both ends to pierce the sealing membrane 22 corresponding to the reserved liquid pack 126, allowing the test solution within the reserved liquid pack 126 to flow naturally into the reserved liquid chamber 126 for temporary storage. The placement and operation of the piercing needle 9 at other reagent pack units are similar.

[0115] In the above embodiments, see Figure 7 Four PCR chambers are arranged side by side, with adjacent PCR chambers staggered vertically, so that the center line connecting each PCR chamber forms a parallelogram, effectively saving space. The PCR chambers are through holes on the cartridge plate 11, and are sealed by the front membrane 12 and the back membrane 13 attached to the front and back of the cartridge plate 11.

[0116] The present invention also provides a detection device, including the above-mentioned card box and a matching instrument used in conjunction with the card box. The accompanying instrument is equipped with a receiving slot to allow the card box to be placed vertically. The accompanying instrument mainly includes: an air pump corresponding to each airflow channel and air inlet; a drive module (center rod) corresponding to each valve, which is used to drive the opening and closing of the needle valve 6; a magnetic attraction module corresponding to the magnetic bead capture chamber 108, which is used to provide magnetic attraction force to control the attraction or release of magnetic beads; an ultrasonic module and a thermal circulation module corresponding to the sample processing chamber 103, which are used to mix and heat the sample mixture; a pressing module corresponding to the piercing needle 9, which is used to provide a predetermined pressing force to squeeze the piercing needle 9 to pierce the sealing film 22 of the reagent package unit; a quantitative detection module corresponding to the first quantitative chamber 110 and the second quantitative chamber 106, which is used to detect the amount of liquid in the first quantitative chamber 110 or the second quantitative chamber 106; and an amplification heating module and an amplification detection module corresponding to the PCR chamber. The amplification heating module is used to cycle the temperature of the PCR chamber, and the amplification detection module is used to detect the amplification status in the PCR chamber. The quantitative detection module can use a high-definition camera to monitor and photograph the liquid surface in real time and then intelligently identify the liquid level to detect the amount of liquid in the cavity; or it can use a pressure sensor to monitor the pressure in the cavity and determine the amount of liquid in the cavity based on the pressure model.

[0117] The usage process of the card holder and detection device proposed in this invention is as follows:

[0118] Step 1, Sample loading: Add the patient's sample to be tested into the sample loading cavity 101 of the cartridge body 1, and then close the cap 3 to seal the cartridge. Since there is a siphon bend 102 between the sample loading cavity 101 and the sample processing cavity 103, the sample will temporarily stay in the siphon bend 102 and will not flow directly into the sample processing cavity 103.

[0119] Step 2: Connect the accessory instrument: Insert the cartridge into the accessory instrument. Positioning pins 7 are located on the cartridge body 1, and the entire cartridge is fixed to the accessory instrument via these pins. All air pump holes on the cartridge body 1 are connected to the air pump of the accessory instrument, and the accessory instrument drives all center rods to close all pin valves 6, thus closing all channels within the cartridge body 1.

[0120] Step 3, Sample Pretreatment: After the detection begins, air is drawn from the air pump port of the eighth airway A8, causing the sample remaining in the siphon bend 102 to be sucked into the sample processing chamber 103. The desiccant stored in the condenser chamber 104 absorbs liquid in the air, preventing sample liquid from being mixed with air and sucked into the eighth airway A8, thus contaminating the instrument. The instrument releases the center rod corresponding to the needle valves B2, B3, and B5, putting the needle valves B2, B3, and B5 in the open state (the operation of the center rod of the instrument will not be mentioned again in the following description, only the on / off state of the needle valves will be described). At the same time, the instrument's air pump introduces air into the sample processing chamber 103 through the seventh airway A7 (the operation of the air pump of the instrument will not be mentioned again in the following description, only the air passage ventilation or extraction state will be described), causing bubbles to continuously form at the bottom of the sample processing chamber 103. The bubbles are used to mix the sample with the pre-stored spotting reagent in the sample processing chamber 103. After the spotting reagent and sample are thoroughly mixed, it helps to digest potential nucleases in the sample. Close the pin valves B2, B3, and B5, and activate the ultrasonic module and thermal circulation module located outside the sample processing chamber 103 to accelerate the mixing and digestion of the sample inside the sample processing chamber 103.

[0121] Step 4: Release the lysis buffer: Drive the puncture needle 9 via the pressing module of the accompanying instrument to puncture the sealing membrane 22 of the lysis buffer package 121, allowing the lysis buffer to flow from the package 121 into the test solution chamber 132 (the operation of the pressing module of the accompanying instrument will not be mentioned again below; only the status of the reagent package will be described). At this time, because the corresponding valve of the channel is closed, the lysis buffer will temporarily remain in the test solution chamber 132 and will not automatically flow into the reaction chambers. Open valves B2, B3, B5, B6, and B11, and introduce air through the first airflow channel A1, allowing the lysis buffer to enter the sample processing chamber 103. By continuously introducing air through the first airflow channel A1, the lysis buffer and sample are thoroughly mixed using the bubbles continuously generated from the bottom of the sample processing chamber 103. Then close valves B2, B3, B5, B6, and B11. During the mixing process, the mixture can be heated to 85°C for thermal lysis via the thermal circulation module. In this step, if a quantitative amount of lysis buffer needs to be measured, it can be achieved through the second quantitative chamber 106. The specific operation is as follows: Open the needle valves B6 and B11, and introduce air through the first airflow channel A1 to allow the lysis buffer to enter the second quantitative chamber 106. At this time, the quantitative detection module of the accompanying instrument detects the liquid level in the chamber to determine the amount of liquid measured. When the required amount of liquid is reached, stop pushing in the lysis buffer and close the needle valves B6 and B11. Then open the needle valves B2, B3, and B5, and introduce air through the seventh airflow channel A7 to push the quantitative lysis buffer in the second quantitative chamber 106 into the sample processing chamber 103. By continuously introducing air through the seventh airflow channel A7, the bubbles continuously generated from the bottom of the sample processing chamber 103 ensure thorough mixing of the lysis buffer and the sample. Then close the needle valves B2, B3, and B5.

[0122] Step 5: Release the binding fluid: Puncture the binding fluid package 122, open the needle valves B1, B5, B6, and B10, and introduce air through the second airflow channel A2. Push the binding fluid into the magnetic bead storage chamber 105 to dissolve the lyophilized magnetic beads, mix it thoroughly, and carry the magnetic beads to the sample processing chamber 103. Continue to introduce air through the second airflow channel A2, using the bubbles continuously generated from the bottom of the sample processing chamber 103 to mix the binding fluid, magnetic beads, and the lysed sample. Then close the needle valves B1, B5, B6, and B10. If a quantitative amount of binding fluid needs to be measured in this step, it can be achieved through the second quantitative chamber 106. The specific operation is as follows: open the needle valves B6 and B10, and introduce air through the second airflow channel A2 to allow the binding fluid to enter the second quantitative chamber 106. At this time, use the quantitative detection module of the accompanying instrument to detect the liquid level in the chamber to determine the amount of liquid measured. When the required amount of liquid is reached, stop pushing in the binding fluid and close the needle valves B6 and B10. Then, open the needle valves B1 and B5, and introduce air through the seventh airway A7 to push the quantitative lysis solution in the second quantitative chamber 106 sequentially into the lyophilized magnetic bead chamber 105 and the sample processing chamber 103. By continuously introducing air through the seventh airway A7, the lysis solution and sample are thoroughly mixed using the bubbles continuously generated from the bottom of the sample processing chamber 103. Then, close the needle valves B1 and B5.

[0123] In the above steps, the lysis buffer is released first, followed by the release of the binding solution and mixing with the magnetic beads, and then they are mixed together. However, in the scheme of this application, the lysis buffer can also be released first and mixed with the magnetic beads, and then the binding solution can be released and mixed together. The specific operation of this method is as follows:

[0124] Pierce the lysis buffer package 121, open the valves B1, B5, B6, and B11, and introduce air through the first airflow channel A1. The lysis buffer is first pushed into the magnetic bead storage chamber 105 to dissolve the lyophilized magnetic beads, mixes them thoroughly, and carries the magnetic beads to the sample processing chamber 103. Continue to introduce air through the first airflow channel A1, using the bubbles continuously generated from the bottom of the sample processing chamber 103 to mix the lysis buffer, magnetic beads, and sample. Then close the valves B1, B5, B6, and B11. Pierce the binding fluid package 122, open the valves B2, B3, B5, B6, and B10, and introduce air through the second airflow channel A2, allowing the binding fluid to enter the sample processing chamber 103. Continue to introduce air through the second airflow channel A2, using the bubbles continuously generated from the bottom of the sample processing chamber 103 to thoroughly mix the binding fluid, magnetic beads, and lysed sample. Then close the valves B2, B3, B5, B6, and B10. The procedure for measuring a fixed amount of lysis solution or binding solution is the same as the previous method, and will not be repeated here.

[0125] Step 6, Magnetic Bead Capture: Open the needle valves B2, B3, and B12 to introduce air through the eighth airway A8. Push the mixed lysis buffer, binding buffer, and magnetic beads from the sample processing chamber 103 into the magnetic bead capture chamber 108. Under the magnetic attraction of the magnetic module outside the magnetic bead capture chamber 108, the magnetic beads will be uniformly adsorbed onto the inner wall surface of the magnetic bead capture chamber 108, while other reagents are pushed into the waste liquid chamber 107. Then close the needle valves B2, B3, and B12.

[0126] Step 7: Release the cleaning solution: Puncture the first cleaning solution pack 123, open the ejector valves B5, B6, B9, and B12, and introduce air through the third airflow channel A3 to push the cleaning solution into the magnetic bead capture chamber 108. After thoroughly washing the magnetic beads containing extracts of biological macromolecules such as nucleic acids that are uniformly adsorbed on the inner wall surface of the magnetic bead capture chamber 108, the solution is pushed into the waste liquid chamber 107. Then, close the ejector valves B5, B6, B9, and B12. For the second cleaning, puncture the second cleaning solution pack 125, open the ejector valves B8, B6, B5, and B12, and introduce air through the fifth airflow channel A5 to push the second cleaning solution into the magnetic bead capture chamber 108. After thoroughly washing the magnetic beads containing extracts of biological macromolecules such as nucleic acids that are uniformly adsorbed on the inner wall surface of the magnetic bead capture chamber 108, the solution is pushed into the waste liquid chamber 107. Then, air is continuously introduced through the fifth airflow channel A5 to dry the reagent remaining on the surface of the magnetic beads in the magnetic bead capture chamber 108, and then the needle valves B8, B6, B5, and B12 are closed. In this step, if a quantitative amount of cleaning solution needs to be measured, it can be achieved through the second quantitative chamber 106. The specific operation is the same as that for measuring a quantitative amount of lysis solution, and will not be described again here.

[0127] Step 8: Release the eluent: Puncture the eluent pack 124, open the valves B2, B4, B13, and B14, and introduce air through the fourth airflow channel A4 to push the eluent into the mixing chamber 109. Continue to introduce air through the fourth airflow channel A4. The bubbles generated at the bottom of the mixing chamber 109 will mix the eluent with the freeze-dried beads in the mixing chamber 109 to form an eluent mixture. Then close the valves B13 and B14.

[0128] Step 9: Measure a certain amount of elution mixture and push it into the PCR chamber: Open valve B15 and introduce air through the eighth airway A8 to push a portion of the elution mixture into the first quantitative chamber 110. Using the detection module of the accompanying instrument, after determining that the predetermined volume (25 μL) of elution mixture has been measured in the first quantitative chamber 110, close valves B2, B4, and B15. Open valve B16 and introduce air through the ninth airway A9 to push the elution mixture collected in the first quantitative chamber 110 into PCR chamber 111, then close valve B16.

[0129] Reopen valves B2, B4, and B15, and introduce air through the eighth airway A8 to push a portion of the elution mixture into the first quantitative chamber 110. Using the detection module of the accompanying instrument, after determining that a predetermined volume (25 μL) of elution mixture has been measured in the first quantitative chamber 110, close valves B2, B4, and B15. Open valve B17, and introduce air through the ninth airway A9 to push the elution mixture collected in the first quantitative chamber 110 into PCR chamber 112 (number 2), then close valve B17.

[0130] Reopen valves B2, B4, and B15, and introduce air through the eighth airway A8 to push a portion of the elution mixture into the first quantitative chamber 110. Using the detection module of the accompanying instrument, after determining that a predetermined volume (25 μL) of elution mixture has been measured in the first quantitative chamber 110, close valves B2, B4, and B15. Open valve B18, and introduce air through the ninth airway A9 to push the elution mixture collected in the first quantitative chamber 110 into PCR chamber 113 (number 3), then close valve B18.

[0131] Reopen valves B2, B4, and B15, and introduce air through the eighth airway A8 to push a portion of the elution mixture into the first quantitative chamber 110. Using the detection module of the accompanying instrument, after determining that a predetermined volume (25 μL) of elution mixture has been measured in the first quantitative chamber 110, close valves B2, B4, and B15. Open valve B19, and introduce air through the ninth airway A9 to push the elution mixture collected in the first quantitative chamber 110 into PCR chamber 4 114, then close valve B19.

[0132] Step 10: Cycle the four PCR chambers to temperature using the amplification heating module of the accompanying instrument, and detect the amplification results using the amplification detection module. After the experiment, use the accompanying instrument to heat-seal the top area (the section of the tenth airway A10 near its vent) and the bottom area (the section of the sixth flow channel 146 located between the second quantitative chamber 110 and the needle valve B16) of the four PCR chambers. This seals the PCR chambers and prevents amplification products from leaking out of the cartridges and contaminating the accompanying instrument and the environment.

[0133] In summary, this invention proposes an integrated microfluidic nucleic acid extraction, amplification, and detection cartridge and detection device. The reaction chambers (such as the sample chamber, magnetic bead storage chamber 105, magnetic bead capture chamber 108, waste liquid chamber 107, mixing chamber 109, and PCR chamber) are rationally arranged on the cartridge body 1. The reagents required for the reaction are independently sealed in a reagent pack also located on the cartridge body 1, thus solving the reagent storage problem. Preferably, a puncture needle structure is used to puncture the reagent pack.

[0134] In addition, the cartridge body 1 is rationally designed with airflow channels, main flow channels and valves (needle valves). The flow direction and stopping position of the sample and reagent are controlled by switching the needle valve on and off. The flow of liquid in the cartridge body 1 is controlled by driving the liquid with an air pump, so that the reagent pack and the reaction chamber are selectively connected, and the control is simple and reliable.

[0135] Furthermore, when using an air pump to control the liquid within the cartridge, or when the liquid reacts within the reaction chamber, bubbles are easily generated. Existing technologies use cartridges with a horizontal structure, making it difficult to remove these bubbles generated during the reaction or when the air pump is applied. In contrast, the cartridge body 1 of this invention has a vertical structure, allowing bubbles generated during the reaction to rise from the bottom up, making them easily eliminated.

[0136] Therefore, the microfluidic nucleic acid extraction, amplification, and detection integrated cartridge and device proposed in this invention can automate the processing of nucleic acid extraction, amplification, and detection, resulting in more accurate detection results. It requires only a few manual operation steps, making it simple, safe, and convenient to use. Using this device, the detection time is less than one hour, with manual operation taking no more than 2 minutes. Furthermore, it can be used to detect pathogen genomic targets in a variety of different human clinical samples, demonstrating wide applicability. The device is stored and transported at room temperature, eliminating the need for a cold chain, thus offering high economic efficiency. The cartridge of this invention is fully enclosed during the detection process, and its detection results are completely consistent with conventional methods. No professional training is required for operation, making it safe and convenient.

[0137] The above description is only a preferred embodiment of the present invention. It should be noted that for those skilled in the art, several improvements and substitutions can be made without departing from the technical principles of the present invention, and these improvements and substitutions should also be considered within the scope of protection of the present invention.

Claims

1. A card holder, characterized in that, The card box is used to vertically insert the matching instrument and includes the card box body (1), reagent pack group (2), cap (3) and waterproof and breathable membrane (4). The card box body (1) is provided with a main channel, a valve group, a reaction chamber and an airflow channel; the valve group includes several valves located at predetermined positions in the main channel, so that one end of the reaction chamber and the airflow channel can be selectively connected to the main channel, and the other end of the airflow channel has a vent for connecting an air pump, and the waterproof and breathable membrane (4) is used to seal the vent. The reaction chamber includes a sample chamber, a magnetic bead storage chamber (105), a magnetic bead capture chamber (108), a waste liquid chamber (107), a mixing chamber (109), a first quantitative chamber (110), and a PCR chamber; the sample chamber is connected to the first end of the magnetic bead storage chamber (105) and the magnetic bead capture chamber (108), respectively; the first end of the magnetic bead capture chamber (108) is also connected to the mixing chamber (109), and its opposite second end is connected to the waste liquid chamber (107); the mixing chamber (109) is connected to the first end of the first quantitative chamber (110), and the opposite second end of the first quantitative chamber (110) is connected to the PCR chamber; the sample chamber has a sample dispensing port (5), and the cap (3) is used to seal the sample dispensing port (5); The reagent pack group (2) is attached to the surface of the card box body (1). The reagent pack group (2) includes at least four independently sealed reagent pack units. The four reagent pack units are a lysis buffer pack (121), a binding buffer pack (122), a first cleaning buffer pack (123), and an elution buffer pack (124). The airflow channels include at least a first airflow channel (A1), a second airflow channel (A2), a third airflow channel (A3), and a fourth airflow channel (A4) corresponding to the four reagent pack units respectively; after the reagent pack unit is broken, the test solution inside flows into the first airflow channel (A1), the second airflow channel (A2), the third airflow channel (A3), and the fourth airflow channel (A4) respectively, 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; The first gas flow channel (A1) corresponds to the lysis buffer package (121), and the second gas flow channel (A2) corresponds to the binding buffer package (122); the main flow channel includes a first branch channel (141) and a second branch channel (142). The two ends of the first branch channel (141) are respectively connected to the bottom end of the sample cavity and the bottom end of the first airflow channel (A1); One end of the second branch channel (142) is connected to the bottom of the sample cavity, and the other end is connected to the first branch channel (141). The connection point is the first connection point (C1). The two ends of the magnetic bead storage cavity (105) are respectively connected to the second branch channel (142). The bottom end of the second airflow channel (A2) is connected to the pipe section on the first branch channel (141) located between the first airflow channel (A1) and the first connection point (C1); The valves are provided at predetermined positions in the first branch channel (141) and the second branch channel (142) to control the lysis solution in the lysis solution package (121) and the binding solution in the binding solution package (122) to flow directly into the sample cavity or flow sequentially through the magnetic bead storage cavity (105) and the sample cavity. The reagent kit assembly (2) includes a shell (21) and a sealing film (22); the shell (21) is provided with at least four liquid-containing chambers (23), each liquid-containing chamber (23) is pre-filled with a corresponding test solution, and a groove (24) is provided at the edge of each liquid-containing chamber (23); the number of sealing films (22) corresponds to the number of liquid-containing chambers (23), and the sealing films (22) are attached to the grooves (24) to seal the liquid-containing chambers (23), thereby forming the reagent kit unit; the valve is a needle valve (6); the needle valve (6) has a channel inside, and a sealing film (61) is provided on the channel; the sealing film (61) deforms under a predetermined pressure, thereby sealing the channel.

2. The card holder according to claim 1, characterized in that, The reagent pack group (2) also includes a second cleaning solution pack (125) and a reserve solution pack (126); the airflow channel also includes a fifth airflow channel (A5) and a sixth airflow channel (A6). The third airflow channel (A3) corresponds to the first cleaning fluid pack (123), and the bottom end of the third airflow channel (A3) is connected to the pipe section on the first branch channel (141) located between the second airflow channel (A2) and the first connection point (C1); The fifth airflow channel (A5) corresponds to the second cleaning fluid pack (125), and the bottom end of the fifth airflow channel (A5) is connected to the pipe section on the first branch channel (141) located between the third airflow channel (A3) and the first connection point (C1); The sixth airflow channel (A6) corresponds to the reserved liquid package (126), and the bottom end of the sixth airflow channel (A6) is connected to the pipe section on the first branch channel (141) located between the fifth airflow channel (A5) and the first connection point (C1).

3. The card holder according to claim 2, characterized in that, The reaction chamber also includes a second metering chamber (106); the top of the second metering chamber (106) is provided with a seventh air passage (A7), and the other end of the seventh air passage (A7) is provided with the air inlet; the bottom of the second metering chamber (106) is connected to a pipe section on the first branch channel (141) located between the sixth air passage (A6) and the first connection point (C1), and the valves are respectively provided on both sides of the connection point.

4. The card holder according to claim 1, characterized in that, The main channel also includes a third branch channel (143), a predetermined section of which forms the magnetic bead capture chamber (108); the first end of the third branch channel (143) is connected to the first connection point (C1), and its opposite second end is connected to one end of the waste liquid chamber (107), and the other end of the waste liquid chamber (107) is provided with an outlet channel. The fourth airflow channel (A4) corresponds to the eluent pack (124), and the bottom end of the fourth airflow channel (A4) is connected to the second end of the third branch channel (143); The valve is provided at a predetermined position in the third branch channel (143) to control the liquid in the magnetic bead capture chamber (108) to flow from the second end of the third branch channel (143) into the waste liquid chamber (107), and to control the eluent in the eluent package (124) to flow from the second end of the third branch channel (143) into the magnetic bead capture chamber (108).

5. The card holder according to claim 1, characterized in that, The sample chamber includes a sample inlet chamber (101) and a sample processing chamber (103). The sample inlet chamber (101) and the sample processing chamber (103) are connected by a siphon bend (102). One end of the siphon bend (102) is connected to the bottom of the sample inlet chamber (101), and the other end is connected to the top of the sample processing chamber (103). The top of the sample inlet chamber (101) is provided with the sample inlet (5). The top of the sample processing chamber (103) is provided with an eighth airway (A8).

6. The card holder according to claim 5, characterized in that, A condensation chamber (104) is provided at a predetermined position in the eighth airway (A8), and a water-absorbing agent is pre-placed in the condensation chamber (104).

7. The card holder according to claim 4, characterized in that, The main channel also includes a fourth branch channel (144), a fifth branch channel (145) and a sixth branch channel (146). One end of the fourth branch channel (144) is connected to the bottom of the sample processing chamber (103), and the other end is connected to the top of the mixing chamber (109). One end of the fifth branch channel (145) is connected to the first end of the third branch channel (143), and the other end is connected to the first end of the mixing chamber (109); One end of the sixth branch channel (146) is connected to the opposite second end of the mixing chamber (109), and the other end is connected to the PCR chamber; the top of the PCR chamber is provided with a tenth airway (A10). The top of the first quantitative chamber (110) is provided with a ninth airway (A9), and the other end of the ninth airway (A9) is provided with the air inlet; the bottom of the first quantitative chamber (110) is connected to the tube segment of the sixth branch channel (146) located between the mixing chamber (109) and the PCR chamber, and the valves are respectively provided on both sides of the connection point.

8. The card holder according to claim 7, characterized in that, Several PCR chambers are arranged side by side, and each PCR chamber has a valve at its bottom that allows it to be selectively connected to the first quantitative chamber (110).

9. The card holder according to claim 1, characterized in that, The airflow channel includes an upper airway (131), a test liquid chamber (132), and a lower flow channel (133); one end of the upper airway (131) has the air inlet, and the other end is connected to the upper part of the test liquid chamber (132); one end of the lower flow channel (133) is connected to the lower part of the test liquid chamber (132), and the other end is connected to the main flow channel; the test liquid chamber (132) corresponds to the liquid-containing chamber (23), and after the reagent pack unit is broken, the test liquid inside it flows into the test liquid chamber (132) and the lower flow channel (133) in sequence.

10. The card holder according to claim 9, characterized in that, A puncture needle (9) is provided in the test liquid chamber (132) at a predetermined distance from the sealing membrane (22).

11. The card holder according to claim 1, characterized in that, The card box body (1) includes a card box plate (11), a front film (12) and a rear film (13). The front side of the cartridge plate (11) is provided with a plurality of recesses, and the front membrane (12) is attached to the front side of the cartridge plate (11) to seal the recesses, thereby forming the other reaction chambers besides the PCR chamber. The card tray plate (11) has several through holes. The front membrane (12) is attached to the front side of the card tray plate (11), and the back membrane (13) is attached to the back side of the card tray plate (11) to seal the through holes, thereby forming the PCR chamber. The card holder plate (11) is provided with a plurality of grooves, and the front film (12) is attached to the front side of the card holder plate (11) to seal the grooves, thereby forming the airflow channel and the main flow channel.

12. A detection device, characterized in that, Includes a card holder and supporting instrument as described in any one of claims 1-11, wherein the supporting instrument includes an air pump communicating with each airflow channel and each air vent, and the supporting instrument is provided with a receiving slot for vertically inserting the card holder into the supporting instrument.

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