CN115386485A - A cartridge and detection device - Google Patents

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, a sealing cover and a waterproof breathable film; 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; 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. 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

A kind of cartridge and detection device

technical field

The invention relates to the field of biomedical technology, in particular to a cartridge and a detection device.

Background technique

Nucleic acid detection plays a very important role in many fields of biochemical analysis and has been widely used in the field of biomedicine.

In the prior art, the spin column method or magnetic bead method is usually used to extract nucleic acid from the sample, which generally requires four steps of lysis, binding, washing, and elution. After the nucleic acid is extracted from the sample, follow-up steps such as hybridization of nucleic acid molecules, polymerase chain reaction (PCR) and biochip are continued to complete nucleic acid detection.

Due to the many steps of nucleic acid detection, it is very difficult to realize the entire "from sample to result" fully automated detection equipment. Moreover, as far as the transfer of active ingredients in each step is concerned, manual transfer is often used in the prior art, which is not only cumbersome, time-consuming and laborious, but also difficult to transfer materials sufficiently and efficiently, and manual operation can easily lead to unstable results. Detection is difficult to implement.

In addition, the mainstream technology of molecular detection is fluorescent quantitative PCR technology. Due to the characteristics of exponential amplification template in PCR technology, the existing open consumables make the whole operation process easy to cause PCR aerosol pollution, affecting the purity of the extract, thus limiting Fluorescent quantitative PCR technology was further applied in clinical practice.

Therefore, prior art needs to be improved urgently.

Contents of the invention

The purpose of the present invention is: the present invention provides a cartridge and a detection device to solve the difficulty in realizing fully automatic processing of nucleic acid detection in the prior art, as well as the instability of detection results caused by manual operation and the use of open consumables. The technical problems of low accuracy and low detection efficiency.

In order to achieve the above object, the present invention provides a cartridge, including a cartridge body, a reagent pack, a cover and a waterproof and breathable membrane;

The cartridge body is provided with a main flow channel, a valve group, a reaction chamber and an air flow channel; the valve group includes several valves arranged at predetermined positions of the main flow channel, so that the reaction chamber and the air flow channel One end of the air channel is selectively communicated with the main channel, the other end of the air channel has a vent for connecting to an air pump, and the waterproof and breathable membrane is used for sealing the vent;

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 respectively connected to the magnetic bead storage chamber and the The first end of the magnetic bead capture chamber is communicated; the first end of the magnetic bead capture chamber is also communicated with the mixing chamber, and its opposite second end is communicated with the waste liquid chamber; the mixing chamber is connected with the The first end of the first quantitative chamber is connected, and the opposite second end of the first quantitative chamber is connected with the PCR chamber; the sample chamber has a sample port, and the cover is used to seal the PCR chamber. Sample port;

The reagent pack set is attached on the surface of the cartridge body, and the reagent pack set includes at least four independently sealed reagent pack units; the four reagent pack units are respectively a lysate pack, a binding solution pack, A first wash solution bag and an eluent bag;

The air channels at least include a first air channel, a second air channel, a third air channel and a fourth air channel respectively corresponding to the four reagent pack units; after the reagent pack unit is broken, its built-in test solution corresponds to flow into the first air channel, the second air channel, the third air channel and the fourth air channel, and flow into one or the other of the reaction chamber along the main channel under the control of the valve group and the air pump Multiple.

In some embodiments of the present application, the first air channel corresponds to the lysate package, and the second air channel corresponds to the binding solution package; the main channel includes a first branch channel and a second branch channel;

Both ends of the first branch flow channel communicate with the bottom end of the sample chamber and the bottom end of the first flow channel respectively;

One end of the second branch flow channel is connected to the bottom end of the sample chamber, and the other end is connected to the first branch flow channel, which is the first connection point, and the two ends of the magnetic bead storage chamber are connected to into the second branch channel;

The bottom end of the second air flow channel is connected to the pipe section on the first branch flow channel between the first air flow channel and the first connection point;

The predetermined position of the first branch flow channel and the second branch flow channel is provided with the valve to control the lysate in the lysate bag and the binding liquid in the binding liquid bag to directly flow into the sample cavity or sequentially Flow through the magnetic bead storage cavity and the sample cavity.

In some embodiments of the present application, the reagent package set further includes a second cleaning solution package and a reserved solution package; the air flow channel also includes a fifth air channel and a sixth air channel;

The third air channel corresponds to the first cleaning liquid bag, and the bottom end of the third air channel is connected to the first branch channel between the second air channel and the first connection point. Pipe section;

The fifth air channel corresponds to the second cleaning liquid bag, and the bottom end of the fifth air channel is connected to the first branch channel between the third air channel and the first connection point. Pipe section;

The sixth air channel corresponds to the reserved liquid bag, and the bottom end of the sixth air channel is connected to a pipe section on the first branch channel between the fifth air channel and the first connection point.

In some embodiments of the present application, the reaction chamber further includes a second quantitative chamber; the top of the second quantitative chamber is provided with a seventh airway, and the other end of the seventh airway has the vent; The bottom of the second quantitative chamber is connected to the pipe section on the first branch flow channel between the sixth flow channel and the first connection point, and the valves are respectively provided on both sides of the connection point.

In some embodiments of the present application, the main flow channel further includes a third branch flow channel, and a predetermined area of the third branch flow channel forms the magnetic bead capture cavity; the first end of the third branch flow channel is connected to the A first connection point, the opposite second end of which communicates with one end of the waste liquid chamber, and the other end of the waste liquid chamber is provided with an air outlet;

The fourth flow channel corresponds to the eluent package, and the bottom end of the fourth flow channel is connected to the second end of the third branch flow channel;

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

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 communicates with the sample processing chamber through a siphon elbow;

One end of the siphon elbow communicates with the bottom of the sample adding chamber, and the other end communicates with the top of the sample processing chamber; the top of the sample adding chamber is provided with the sample port; the top of the sample processing chamber There is an eighth airway.

In some embodiments of the present application, a condensation chamber is provided at a predetermined position of the eighth air passage, and a water-absorbing agent is preset in the condensation chamber.

In some embodiments of the present application, the main channel further includes a fourth branch channel, a fifth branch channel and a sixth branch channel;

One end of the fourth branch channel communicates with the bottom of the sample processing chamber, and the other end communicates with the top of the mixing chamber;

One end of the fifth branch channel communicates with the first end of the third branch channel, and the other end communicates with the first end of the mixing chamber;

One end of the sixth branch channel communicates with the opposite second end of the mixing chamber, and the other end communicates with the PCR chamber; the top of the PCR chamber is provided with a tenth airway;

The top of the first quantitative chamber is provided with a ninth airway, and the other end of the ninth airway has the vent; the bottom of the first quantitative chamber is connected to the sixth branch channel and is located at the The pipe section between the mixing chamber and the PCR chamber, and the valves are respectively provided on both sides of the access.

In some embodiments of the present application, several PCR chambers are provided side by side, and the bottom of each PCR chamber is provided with the valve so that it can be selectively communicated with the first quantitative chamber.

In some embodiments of the present application, the valve is a thimble valve; the thimble valve has a channel inside, and a sealing film is provided on the channel, and the sealing film deforms when subjected to a predetermined pressure, thereby closing the channel.

In some embodiments of the present application, the reagent package set includes a housing and a sealing film; the housing is provided with at least four liquid-containing cavities, and corresponding test solutions are preset in the liquid-containing cavities, and the liquid-containing cavities The edge of the cavity is provided with a circle of sticking grooves; the number of the sealing film corresponds to the number of the liquid-holding chamber, and the sealing film is attached to the sticking groove to seal the liquid-holding cavity, thereby forming the Reagent pack unit.

In some embodiments of the present application, the air flow channel includes an upper air channel, a test solution chamber and a lower flow channel; one end of the upper air channel has the vent, and the other end communicates with the upper part of the test solution chamber; One end of the lower flow channel communicates with the lower part of the test solution chamber, and the other end is connected to the main channel; the test solution chamber corresponds to the liquid containing chamber, and the reagent pack unit has a built-in test solution after it is broken. sequentially flow into the test solution chamber and the lower flow channel.

In some embodiments of the present application, a puncture needle is arranged in the test solution chamber at a predetermined distance from the sealing film.

In some embodiments of the present application, the cartridge body includes a cartridge plate, a front film and a rear film;

The front of the cartridge plate is provided with several concave cavities, and the front film is pasted on the front of the cassette plate to seal the concave cavities, thereby forming other reaction chambers except the PCR cavity. Cavity;

The cartridge board is provided with several through holes, the front film is pasted on the front side of the cartridge board, and the rear film is pasted on the back side of the cartridge board to seal the through holes , thereby forming the PCR chamber;

The cartridge plate (11) is provided with several grooves, and the front film is pasted on the front surface of the cartridge plate to seal the grooves, thereby forming the air flow channel and the main flow channel.

The present application also proposes a detection device, which includes the cartridge described in any one of the above items and supporting instruments.

Compared with the prior art, a cartridge and a detection device according to the embodiment of the present invention have the following beneficial effects:

In the cartridge and detection device of the embodiment of the present invention, each reaction chamber is arranged on the cartridge body, and the reagents required for the reaction are independently sealed in the reagent package group attached to the cartridge body, and the supporting equipment is controlled in the The main channel, air channel and valve on the cartridge body make the reagent package and the reaction chamber selectively communicated, so that the automatic processing of nucleic acid extraction, amplification and detection can be realized. Only a few manual steps are required, and the operation is simple and safe. Convenient, strong sealing, and more accurate detection results.

Description of drawings

In order to more clearly illustrate the technical solutions in the embodiments of the present invention, the following will briefly introduce the accompanying drawings that need to be used in the descriptions of the embodiments or the prior art. Obviously, the accompanying drawings in the following description are only of the present invention. For some embodiments, those of ordinary skill in the art can also obtain other drawings based on these drawings without any creative effort.

Fig. 1 is the exploded structure diagram of the cartridge of the embodiment of the present invention;

Figure 2 is a schematic diagram of the front structure of the cartridge and the central rod of the supporting instrument;

Fig. 3 is a schematic view of the front structure of the cartridge with the front film removed;

Fig. 4 is a schematic diagram of the back structure of the cartridge;

Fig. 5 is a side view structural schematic diagram of the cartridge;

Fig. 6 is a top view structural schematic diagram of the cartridge;

Fig. 7 is a schematic diagram of the axial side structure of the cartridge plate;

Fig. 8 is a schematic diagram of the front structure of the cartridge board;

Fig. 9 is a schematic diagram of the back structure of the cartridge board;

Fig. 10 is a sectional view of D-D in Fig. 2;

Figure 11 is a schematic diagram of the state of cooperation between the center rod and the thimble valve;

Figure 12 is a second schematic diagram of the cooperation state between the central rod and the thimble valve;

Fig. 13 is a schematic diagram of the axial side structure of the housing;

Fig. 14 is a front structural schematic view of the housing;

Figure 15 is a schematic side view of the housing;

Fig. 16 is a schematic top view of the housing;

Figure 17 is a sectional view of E-E in Figure 3;

Figure 18 is an enlarged view of E1 in Figure 17;

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

In the figure, 1. Cartridge body; 11. Cartridge board; 12. Front film; 13. Back film; 2. Reagent pack; 21. Shell; 22. Sealing film; 23. Liquid chamber; 24. Sticking groove ;3. Covering; 4. Waterproof and breathable membrane; 5. Sample injection port; 6. Thimble valve; 61. Sealing film; 7. Locating pin; 8. Center rod;

101, sample adding chamber; 102, siphon elbow; 103, sample processing chamber; 104, condensation chamber; 105, magnetic bead storage chamber; 106, second quantitative chamber; 107, waste liquid chamber; 108, magnetic bead capture chamber; 109, mixing chamber; 110, the first quantitative chamber; 111, No. 1 PCR chamber; 112, No. 2 PCR chamber; 113, No. 3 PCR chamber; 114, No. 4 PCR chamber;

121. Lysis solution package; 122. Binding solution package; 123. First cleaning solution package; 124. Eluent solution package; 125. Second cleaning solution package; 126. Reserve solution package;

A1, the first air channel; A2, the second air channel; A3, the third air channel; A4, the fourth air channel; A5, the fifth air channel; A6, the sixth air channel; A7, the seventh air channel; A8 , the eighth airway; A9, the ninth airway; A10, the tenth airway;

131, upper airway; 132, test solution cavity; 133, lower flow channel;

141, the first branch channel; 142, the second branch channel; 143, the third branch channel; 144, the fourth branch channel; 145, the fifth branch channel; 146, the sixth branch channel;

1501, air channel; 1502, liquid channel; 151, No. 1 channel; 152, No. 2 channel; 153, No. 3 channel; 154, No. 4 channel.

Detailed ways

The specific implementation manners of the present invention will be further described in detail below in conjunction with the accompanying drawings and embodiments. The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention.

In the description of the present invention, it should be understood that the orientation or positional relationship indicated by the terms "upper", "lower", "left", "right", "top", "bottom" etc. Orientation or positional relationship is only for the convenience of describing the present invention and simplifying the description, and does not indicate or imply that the referred device or element must have a specific orientation, be constructed and operated in a specific orientation, and thus should not be construed as a limitation of the present invention.

It is to be understood that the terms "length", "width", "top", "bottom", "front", "rear", "left", "right", "vertical", "horizontal", "top" , "bottom", "inner", "outer" and other indicated orientations or positional relationships are based on the orientations or positional relationships shown in the drawings, and are only for the convenience of describing the present invention and simplifying the description, rather than indicating or implying No device or element must have a specific orientation, be constructed, and operate in a specific orientation and therefore should not be construed as limiting the invention.

In addition, the terms "first" and "second" are used for descriptive purposes only, and cannot be interpreted as indicating or implying relative importance or implicitly specifying the quantity of indicated technical features. Thus, a feature defined as "first" and "second" may explicitly or implicitly include one or more of these features. In the description of the present invention, "plurality" means two or more, unless otherwise specifically defined.

Referring to Figures 1-6, it is a cartridge according to a preferred embodiment of the present invention, including a cartridge body 1, a reagent pack 2, a cover 3 (rubber plug cover), a waterproof breathable membrane 4 and a positioning pin 7.

The cartridge body 1 is provided with a main channel, a valve group, a reaction chamber and an air channel. The valve group includes several valves arranged at predetermined positions of the main flow channel, so that one end of the reaction chamber and the air flow channel can be selectively communicated with the main flow channel. The other end of the air channel has an air vent for connecting an air pump. The waterproof gas-permeable membrane 4 is used to seal the vent hole by heat-compression welding technology.

The reaction chamber at least 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 communicates with the first ends 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 communicated with the mixing chamber 109 , and its opposite second end is communicated with the waste liquid chamber 107 . The mixing chamber 109 communicates with the first end of the first quantitative chamber 110 , and the opposite second end of the first quantitative chamber 110 communicates with the PCR chamber. The sample cavity has a sample inlet 5, and the cover 3 is used to seal the sample inlet 5.

The reagent pack set 2 is pasted on the surface of the cartridge body 1, and the reagent pack set 2 includes at least four independently sealed reagent pack units. The four reagent pack units are a lysing solution pack 121 , a binding solution pack 122 , a first cleaning solution pack 123 and an eluent pack 124 .

The air channels at least include a first air channel A1 , a second air channel A2 , a third air channel A3 and a fourth air channel A4 respectively corresponding to four reagent package units. After the reagent package unit is broken, its built-in test solution flows into the first air channel A1, the second air channel A2, the third air channel A3 and the fourth air channel A4, and flows into the reaction channel along the main channel under the control of the valve group and the air pump. One or more of the cavities.

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

Referring to FIGS. 1-6 , the cartridge body 1 includes a cartridge plate 11 , a front film 12 and a rear film 13 . The front of the cartridge plate 11 is provided with a cavity, a through hole or a groove, the front film 12 is attached to the front of the cartridge plate 11, and the rear film 13 is attached to the back of the cartridge plate 11 to seal the cavity , through holes or grooves, thereby forming the reaction chambers, main channels and air channels. Specifically, the front film 12 and the rear film 13 can be made of polypropylene plastic films, which are pasted on the surface of the cartridge board 11 by laser welding. In the following description, the orientation descriptions related to left, right, up and down are all based on the front of the cartridge board 11 .

Referring to Figures 7-9, the upper right of the cartridge plate 11 is provided with a sample adding chamber 101, a siphon elbow 102, a sample processing chamber 103, a condensation chamber 104, a magnetic bead storage chamber 105 and a second quantitative chamber 106; There is a mixing chamber 109, a first quantification chamber 110 and four PCR chambers, respectively No. 1 PCR chamber 111, No. 2 PCR chamber 112, No. 3 PCR chamber 113 and No. 4 PCR chamber 114; A reagent pack set 2 is provided; a waste liquid chamber 107 and a magnetic bead capture chamber 108 are provided at the lower left. Magnetic particles such as magnetic beads are preset in the magnetic bead storage chamber 105, such as freeze-dried magnetic beads. The main purpose of making the sample pass through the magnetic bead storage chamber 105 is to extract nucleic acid and other biomacromolecular extracts by using magnetic particles including magnetic beads. , is a common method for nucleic acid extraction in the prior art. Freeze-dried beads are preset in the mixing chamber 109 for mixing with samples and eluents. Freeze-dried beads routinely used in nucleic acid detection in the prior art can be used. The sample processing chamber 103 is preset with a dry spot reagent, and after the dry spot reagent is fully mixed with the sample, it helps to digest potential nucleases in the sample. The dry-spotting reagent of the present application can adopt the dry-spotting reagent conventionally used in nucleic acid detection in the prior art, which can realize the above-mentioned functions.

Wherein, the sample adding chamber 101 is located at the top right of the cartridge plate 11, one end of the siphon elbow 102 communicates with the bottom of the sample adding chamber 101, and the other end communicates with the top of the sample processing chamber 103, and the siphon elbow 102 has two downward bends and two upper bends. Further, some dry reagents are provided in the sample processing chamber 103 . The top of the sample processing chamber 103 is provided with an eighth air channel A8, and a condensation chamber 104 is provided at a predetermined position of the eighth air channel A8. The condensation chamber 104 is located on the left side of the sample processing chamber 103, and a water-absorbing agent is preset in the condensation chamber 104, which can absorb water vapor in the sample.

Those skilled in the art should understand that the specific positions of the above reaction chambers are only a preferred embodiment and not the only choice.

Referring to Fig. 3-6, the reagent pack set 2 is pasted on the upper left back of the cartridge board 11, and the reagent pack set 2 includes six independently sealed reagent pack units, namely: eluent pack 124, lysate pack 121 , the binding solution package 122, the first cleaning solution package 123, the second cleaning solution package 125 and the reserved solution package 126 (the number in the figure only represents the position of the reagent package unit, and does not represent the complete reagent package unit structure). The top left of the cartridge plate 11 is provided with an eluent package 124, a lysate package 121, a binding solution package 122, a first cleaning solution package 123, a second cleaning solution package 125, and a reserved solution package 126 in order from left to right. The fourth air channel A4 , the first air channel A1 , the second air channel A2 , the third air channel A3 , the fifth air channel A5 , and the sixth air channel A6 correspond one-to-one. One end port of the air passage is an air pump hole for ventilation, and the air pump hole is connected with an external air pump. In the drawings of the present application, for the convenience of understanding and labeling, the labeling line is marked at the air pump hole of the airway. For example, the marked line A1 in the figure is marked at the air pump hole of the first airway A1, and the same is true for other airways.

Those skilled in the art should understand that there is a one-to-one correspondence between the reagent pack units and the air channels, but the above arrangement order of the reagent pack units is only a preferred embodiment, not the only choice.

Referring to FIG. 8 , the front side of the cartridge plate 11 is slotted and sealed by the front film 12 to form a main channel, which is composed of several branch channels.

Both ends of the first branch flow channel 141 communicate with the bottom end of the sample processing chamber 103 and the bottom end of the first air flow channel A1 respectively, and the bottom end of the first air flow channel A1 is provided with a valve B11 .

One end of the second branch flow channel 142 is connected to the bottom end of the sample processing chamber 103 , and the other end is connected to the first branch flow channel 141 , and the connection point is the first connection point C1 . Both ends of the magnetic bead storage chamber 105 are respectively connected to the second branch channel 142, and the pipe section between the magnetic bead storage chamber 105 and the sample processing chamber 103 is provided with a valve B1.

The bottom end of the second air channel A2 is connected to the pipe segment between the first air channel A1 and the first connection point C1 on the first branch channel 141 through the valve B10 .

The bottom end of the third air channel A3 is connected to the pipe segment between the second air channel A2 and the first connection point C1 on the first branch channel 141 through the valve B9.

The bottom end of the fifth air channel A5 is connected to the pipe segment between the third air channel A3 and the first connection point C1 on the first branch channel 141 through the valve B8.

The bottom end of the sixth air passage A6 is connected to the pipe section between the fifth air passage A5 and the first connection point C1 on the first branch flow passage 141 through the needle valve.

The top of the second quantitative chamber 106 is provided with a seventh airway A7, and the other end of the seventh airway A7 has a vent for connecting to an air pump. The bottom of the second quantitative chamber 106 is connected to the pipe section on the first branch channel 141 between the sixth air channel A6 and the first connection point C1, and the left side of the connection is provided with a valve B6, and the right side is provided with a valve B5.

A predetermined area of the third branch flow channel 143 forms the magnetic bead capture cavity 108, the first end of the third branch flow channel 143 is connected to the first connection point C1, and its opposite second end communicates with one end of the waste liquid chamber 107, and A first valve B12 is provided at the communicating 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 air channel A4.

The bottom end of the fourth flow channel A4 is connected to the second end of the third branch flow channel 143 through the first valve B13.

One end of the fourth branch flow channel 144 communicates with the top of the mixing chamber 109, and the other end is connected to the first branch flow channel 141, which is the second access point C2, and is connected to the second connection point C2 at the bottom of the sample processing chamber 103. A valve B2 is set between the entry point C2, a valve B3 is set between the first access point C1 and the second access point C2, and a valve B4 is set between the second access point C2 and the mixing chamber 109, so that the fourth The other end of the branch channel 144 can communicate with the bottom of the sample processing chamber 103 through the tube section of the first branch channel 141 .

One end of the fifth branch channel 145 communicates with the first end of the third branch channel 143 , and the other end communicates with the first end of the mixing chamber 109 , and the fifth branch channel 145 is provided with a valve B14 .

One end of the sixth branch channel 146 communicates with the opposite second end of the mixing chamber 109, and the other end communicates with the PCR chamber.

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 an air vent. The top of the PCR cavity is provided with a tenth airway A10, and the other end of the tenth airway A10 has an air vent.

Referring to FIG. 19 , in this embodiment, there are four PCR chambers in total, which are No. 1 PCR chamber 111 , No. 2 PCR chamber 112 , No. 3 PCR chamber 113 and No. 4 PCR chamber 114 . Each PCR cavity corresponds to a channel, the No. 1 PCR cavity 111 corresponds to the No. 1 channel 151, the No. 2 PCR cavity 112 corresponds to the No. 2 channel 152, the No. 3 PCR cavity 113 corresponds to the No. 3 channel 153, and the No. 4 PCR cavity 114 corresponds to No. 4 Channel 154. Specifically, taking the No. 4 channel 154 as an example, the above-mentioned channels all include an air channel 1501 and a liquid channel 1502 . One end of the air channel 1501 communicates with the top of the No. 4 PCR chamber 114, and the other end communicates with the tenth air channel A10. One end of the liquid channel 1502 communicates with the bottom of the No. 4 PCR chamber 114 , and the other end communicates with the sixth branch flow channel 146 .

That is, in this embodiment, the other end of the sixth branch channel 146 communicates with the No. 4 channel 154, and the first quantitative chamber 110, the No. 1 channel 151, the No. 2 channel 152, and the No. 3 channel 153 can all be regarded as connected to the No. 4 channel. Six branch flow channels 146, and the sixth branch flow channel 146 is provided with a valve B15 at a position between the mixing chamber 109 and the first quantitative chamber 110, No. 1 channel 151, No. 2 channel 152, No. 3 channel 153 and 4 Valve B16, valve B17, valve B18 and valve B19 are respectively provided on the liquid channel 1502 of No. channel 154. The air channel 1501 of No. 1 channel 151, No. 2 channel 152, No. 3 channel 153 and No. 4 channel 154 are connected to the Ten Airway A10.

In the above-described embodiment, the cartridge plate 11 is provided with four circular through holes of the same size, and the front film 12 and the rear film 13 are respectively pasted on the front and back of the cartridge plate 11 to seal the through holes, thereby No. 1 PCR chamber 111 , No. 2 PCR chamber 112 , No. 3 PCR chamber 113 and No. 4 PCR chamber 114 are formed. It can be seen from the figure that the four PCR cavities are arranged side by side, and the adjacent PCR cavities are arranged staggered up and down, so that the angle between the line connecting the centers of the four PCR cavities and the horizontal line is 30°-60°, preferably 40°-50° , and form a parallelogram, the above design effectively saves the layout space of the PCR chamber.

In the above embodiments, the volume of the first quantitative chamber 110 has a certain relationship with the volume of the PCR chamber, and the volume of the mixing chamber 109 also has a certain relationship with the total volume of all PCR chambers. Because there will be a certain loss of liquid when passing through the flow channel and the channel, it is preferable to set the volume of the first quantification chamber 110 slightly larger than the volume of the PCR chamber, so that enough eluent mixture can be measured at one time to fill the PCR chamber. Preferably, the ratio of the volume of the first quantification chamber 110 to the volume of the PCR chamber is between 1.1-1.5, so as to reduce the volume ratio of the first quantification chamber 110 on the cartridge plate 11 while meeting the above-mentioned measuring requirements . In addition, 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 be able to store enough elution mixture at one time to fill all the PCR chambers. Preferably, the ratio of the volume of the mixing chamber 109 to the total volume of all PCR chambers is between 1.1-1.5, so as to reduce the volume ratio of the mixing chamber 109 on the cartridge plate 11 while meeting the above storage requirements.

In the above embodiments, all valves B1-B19 are thimble valves 6 . Referring to FIG. 10 , the supporting instrument is provided with a drive module (central rod) 8 to cooperate with the thimble valve 6 . Referring to 11-12, the thimble valve 6 has a channel inside, and a sealing film 61 is arranged on the channel. There is a certain distance between the driving module 8 and the thimble valve 6 in the original state, the sealing film 61 is not under pressure, does not deform, and the channel is unblocked, that is, the thimble valve 6 is in an open state, as shown in FIG. 11 . When the driving module 8 moves forward to fit the thimble valve 6 , the sealing film 61 is deformed under pressure, thereby closing the channel, that is, the thimble valve 6 is in a closed state, as shown in FIG. 12 .

In the above embodiment, referring to FIG. 1, the reagent pack set 2 includes a housing 21 and a sealing film 22. The reagent pack set 2 is integrally arranged on the upper left side of the cartridge plate 11. Specifically, the housing 21 is attached to the cartridge by laser welding. surface of the plate 11. Referring to Figures 13-16, the housing 21 is provided with six liquid chambers 23, and corresponding test liquids are preset in the liquid chambers 23, and the edges of the liquid chambers 23 are provided with sticking grooves 24. The number of sealing films 22 is the same as the number of liquid-holding chambers 23 , and they are attached to the attachment grooves 24 of each liquid-holding chamber 23 in a one-to-one correspondence to seal each liquid-holding chamber 23 to form independent reagent pack units. The sealing film 22 can be an aluminum-plastic film, and each liquid-holding cavity 23 is sealed by a thermocompression welding technique.

The arrangement of the liquid chambers 23 on the housing 21 is as follows: three rows of liquid chambers 23 are arranged from top to bottom, and the liquid chambers 23 in each row are arranged at equal intervals, and each liquid chamber 23 in an even-numbered row is respectively connected to The gaps between every two adjacent liquid chambers 23 in odd rows correspond to each other. The arrangement of the above-mentioned liquid-holding chambers 23 facilitates the reserved space for each liquid-holding chamber 23 to be correspondingly provided with an air passage, so that the arrangement of the air passages is more orderly.

According to the above-mentioned arrangement rules, when the reagent pack group 2 is provided with six reagent pack units in total, see Figure 14 (in Figure 14, each reagent pack unit is marked with the position of the liquid chamber 23), including the lysates arranged in sequence in the first row Package 121, the first cleaning solution package 123 and the reserved solution package 126; the binding solution package 122 between the lysing solution package 121 and the first cleaning solution package 123 on the second row is located between the first cleaning solution package 123 and the reserved solution package. The second cleaning solution package 125 between the solution packages 126; the eluent package 124 corresponding to the position of the lysing solution package 121 on the third row.

The shape of the liquid-holding chamber 23 is diamond-shaped, and its two ends respectively form pointed arcs protruding upward and downward. The depth of each liquid-holding chamber 23 can be set according to the preset test liquid volume, and the ones located in the first row and the first column The depth of the liquid chamber 23 is greater than that of the liquid chambers 23 at other positions, and in this embodiment, the depth of the lysate package 121 is the largest.

The prior art generally only cleans once, but in this application, because the structure of the reagent pack group 2 is adopted, it is more convenient to increase the reagent pack unit, so the second cleaning solution pack 125 is provided to perform secondary cleaning, so that the cleaning effect of the magnetic beads is better. it is good. In addition, the reagent pack set 2 is also provided with a reserved solution pack 126 to reserve space for subsequent technical development or process improvement.

In the above embodiment, referring to FIG. 17 and FIG. 18 , the sixth air channel A6 is taken as an example, and the air channel includes an upper air channel 131 , a test solution chamber 132 and a lower flow channel 133 . One end of the upper airway 131 has an air vent, and the other end communicates with the upper part of the test solution chamber 132 . One end of the lower flow channel 133 communicates with the lower part of the test solution chamber 132, and the other end is connected to the main flow channel. The test solution chamber 132 corresponds to the liquid containing chamber 23 , and after the reagent pack unit is broken, the built-in test solution flows into the test solution chamber 132 and the lower flow channel 133 sequentially, and then flows into the main flow channel.

In the above-described embodiment, a puncture needle 9 is provided at a predetermined distance from the sealing film 22 in the test solution chamber 132, and the puncture needle 9 punctures the sealing film 22 after receiving a predetermined pressing force, so that the test solution in the reagent pack unit flows into the test solution chamber 132. Test solution chamber 132 . Further, the piercing needle 9 is an elastically deformable part, and its two ends are respectively provided with sharp needles 91 facing the sealing film 22, and the pressing module of the matching instrument simultaneously squeezes the sharp needles 91 at both ends to pierce the reserved liquid bag 126 Corresponding to the sealing film 22, the test solution in the reserved liquid bag 126 will naturally flow into the reserved liquid cavity 126 for temporary storage. The setting position and working process of the puncture needle 9 at other reagent pack units are the same.

In the above embodiment, referring to FIG. 7 , four PCR chambers are arranged side by side, and adjacent PCR chambers are staggered up and down, so that the connecting lines between the centers of each PCR chamber form a parallelogram, which effectively saves the layout space. The PCR cavity is a through hole arranged on the cartridge plate 11, and is sealed by a front film 12 and a rear film 13 attached to the front and back of the cartridge plate 11.

The present invention also provides a detection device, including the above-mentioned cartridge, and supporting instruments used in conjunction with the cartridge. The supporting equipment is provided with accommodating slots to allow the cartridges to be placed vertically. The supporting equipment mainly includes: air pumps are provided corresponding to each air channel and the air outlet of the air channel; drive modules are provided corresponding to each valve (central rod), which is used to drive the opening and closing of the thimble valve 6; the position corresponding to the magnetic bead capture chamber 108 is provided with a magnetic attraction module, which is used to provide magnetic attraction force, control the attraction or release of magnetic beads; and the sample processing chamber 103 The corresponding position is provided with an ultrasonic module and a thermal cycle module, which are used to mix and heat the sample mixture; a pressing module is provided at a position corresponding to the puncture needle 9, which is used to provide a predetermined pressing force and squeeze the puncture needle 9 puncture the sealing film 22 of the reagent pack unit; a quantitative detection module is provided at the corresponding position of the first quantitative chamber 110 and the second quantitative chamber 106, which is used to detect The amount of liquid; the corresponding position of the PCR chamber is equipped with an amplification heating module and an amplification detection module. The amplification heating module is used to heat up and cool down the PCR chamber, and the amplification detection module is used to detect the amplification in the PCR chamber. . Among them, the quantitative detection module can use a high-definition camera to monitor the liquid level in real time and take pictures to intelligently identify the liquid level to detect the amount of liquid in the cavity; or use a pressure sensor to monitor the pressure in the cavity and judge the liquid in the cavity according to the pressure model quantity.

The use process of a kind of cartridge and detection device proposed by the present invention is as follows:

The first step, sample loading: add the patient sample to be tested into the sample adding chamber 101 of the cartridge body 1, and then buckle the cover 3 to seal the cartridge. Since the siphon elbow 102 is provided between the sample adding chamber 101 and the sample processing chamber 103 , the sample will temporarily stay in the siphon elbow 102 instead of directly flowing into the sample processing chamber 103 .

The second step, connect the supporting instrument: insert the cartridge into the supporting instrument. The positioning pin 7 is arranged on the cartridge body 1, and the whole cartridge is fixed on the matching instrument through the positioning pin 7. The air pump holes of all the air passages on the cartridge body 1 are connected to the air pumps of the supporting instruments, and the supporting instruments drive all the central rods to close all the thimble valves 6, so that all the channels in the cartridge body 1 are in a closed state.

The third step, sample pretreatment: after the test starts, air is sucked from the air pump hole of the eighth airway A8, so that the sample staying in the siphon elbow 102 is sucked into the sample processing chamber 103 . The water-absorbing agent in the condensation chamber 104 will absorb the liquid in the air, so as to prevent the sample liquid from being mixed in the air and sucked into the eighth airway A8, thus polluting the supporting instruments. Let the supporting instrument release the center rod corresponding to the thimble valve B2, B3, B5, so that the thimble valve B2, B3, B5 is in the open state (the following description does not refer to the operation of the center rod of the supporting instrument, only the switch state of the thimble valve) . At the same time, air is introduced into the sample processing chamber 103 through the seventh airway A7 using the pneumatic pump of the supporting instrument (the operation of the pneumatic pump of the supporting instrument is no longer mentioned in the following description, only the ventilation or pumping state of the airway is explained), so that the sample can be processed Bubbles are continuously generated at the bottom of the chamber 103 , and the sample is mixed with the spot-dried reagent pre-stored in the sample processing chamber 103 by using the air bubbles. After the spot dry reagent is thoroughly mixed with the sample, it helps to digest potential nucleases in the sample. Close the thimble valves B2, B3, B5, and start the ultrasonic module and the thermal cycle module arranged outside the sample processing chamber 103 to accelerate the mixing and digestion of the samples in the sample processing chamber 103.

The fourth step, releasing the lysate: the puncture needle 9 is driven by the pressing module of the supporting instrument, and the sealing film 22 of the lysate bag 121 is pierced, so that the lysate flows out from the lysate bag 121 to the test solution chamber 132 (described below) The operation of the pressing module of the supporting instrument is no longer mentioned, and only the status of the reagent pack is described). At this time, since the thimble valve corresponding to the channel is in a closed state, the lysate will temporarily stay in the test solution chamber 132 and will not automatically flow into each reaction chamber. Open the thimble valves B2, B3, B5, B6, and B11, and pass air through the first air channel A1, so that the lysate enters the sample processing chamber 103 . By continuing to supply air from the first air channel A1, the lysate is fully mixed with the sample by using the air bubbles continuously generated from the bottom of the sample processing chamber 103, and then the needle valves B2, B3, B5, B6, and B11 are closed. During the mixing process, the mixed solution can be heated to 85°C through the thermal cycle module for thermal cracking. In this step, if a quantitative amount of lysate needs to be measured, it can be realized through the second quantitative chamber 106. The specific operation is: open the thimble valves B6 and B11, and pass air through the first air channel A1, so that the lysate enters the second quantitative chamber. In the second quantitative chamber 106. At this time, use the quantitative detection module of the supporting instrument to detect the liquid level in the chamber to judge the amount of liquid measured in the chamber. When the liquid amount reaches the required amount, stop pushing the lysate and close the thimble valves B6 and B11. Then the thimble valves B2, B3, B5 are opened, and air is introduced from the seventh air channel A7 to push the quantitative lysate in the second quantitative chamber 106 into the sample processing chamber 103 . By continuously feeding air from the seventh air channel A7, the lysate and the sample are fully mixed by using the bubbles continuously generated from the bottom of the sample processing chamber 103, and then the thimble valves B2, B3, and B5 are closed.

Step 5. Release the binding solution: puncture the binding solution package 122, open the thimble valves B1, B5, B6, and B10, and let in air from the second air channel A2, and push the binding solution into the magnetic bead storage chamber 105 to dissolve the frozen beads. Dry the magnetic beads, mix and carry the magnetic beads, and finally reach the sample processing chamber 103 . Continue to introduce air from the second air channel A2, use the bubbles continuously generated from the bottom of the sample processing chamber 103 to mix the binding liquid, magnetic beads and the lysed sample, and then close the thimble valves B1, B5, B6, and B10. In this step, if it is necessary to measure a certain amount of binding solution, it can be realized through the second quantitative chamber 106. The specific operation is: open the thimble valves B6 and B10, and pass air through the second air channel A2, so that the binding solution enters the first In the second quantitative chamber 106. At this time, use the quantitative detection module of the supporting instrument to detect the liquid level in the cavity to judge the amount of liquid measured in the cavity. When the liquid amount reaches the required amount, stop pushing the binding solution and close the thimble valves B6 and B10. Then the thimble valves B1 and B5 are opened, and air is introduced from the seventh airway A7 to push the quantitative lysate in the second quantitative chamber 106 into the freeze-dried magnetic bead chamber 105 and the sample processing chamber 103 in sequence. By continuing to supply air from the seventh air channel A7, the lysate and the sample are fully mixed by using the air bubbles continuously generated from the bottom of the sample processing chamber 103, and then the thimble valves B1 and B5 are closed.

In the above steps, the lysate is released first, and then the binding solution is released to mix with the magnetic beads, and then mixed together. However, in the scheme of this application, it is also possible to release the lysate and mix with the magnetic beads first, and then release the binding solution to mix together. The specific operation of this method is as follows:

Pierce the lysate package 121, open the thimble valves B1, B5, B6, and B11, and introduce air from the first air channel A1, push the lysate into the magnetic bead storage chamber 105 to dissolve the freeze-dried magnetic beads, and mix well to carry the magnetic beads. The beads finally reach the sample processing chamber 103. Continue to supply air from the first air channel A1, and mix the lysate, magnetic beads and samples by using the air bubbles continuously generated from the bottom of the sample processing chamber 103, and then close the thimble valves B1, B5, B6, and B11. Puncture the binding solution package 122 , open the thimble valves B2 , B3 , B5 , B6 , and B10 , and let air flow through the second air channel A2 to allow the binding solution to enter the sample processing chamber 103 . By continuously feeding air from the second air channel A2, using the bubbles continuously generated from the bottom of the sample processing chamber 103, the binding solution, magnetic beads and the lysed sample are fully mixed, and then the thimble valves B2, B3, B5, and B6 are closed , B10. The operation of measuring a quantitative lysate or binding solution is the same as the previous method, and will not be repeated here.

Step 6, magnetic bead capture: open the thimble valves B2, B3, B12, let air in from the eighth airway A8, and push the mixed lysate, binding solution and magnetic beads in the sample processing chamber 103 first through the magnetic beads Capture chamber 108, under the action of the magnetic attraction force of the magnetic attraction module outside the magnetic bead capture chamber 108, the magnetic beads will be evenly adsorbed on the inner wall surface of the magnetic bead capture chamber 108, while other reagents are pushed into the waste liquid chamber 107 , and then close the thimble valves B2, B3, B12.

The seventh step, releasing the cleaning solution: puncture the first cleaning solution package 123, open the thimble valves B5, B6, B9, and B12, pass air through the third air channel A3, and push the cleaning solution into the magnetic bead capture chamber 108 first, The magnetic beads with nucleic acid and other biological macromolecular extracts evenly adsorbed on the inner wall surface of the magnetic bead capture chamber 108 are fully washed and then pushed into the waste liquid chamber 107, and then the thimble valves B5, B6, B9, and B12 are closed. When performing the second cleaning, puncture the second cleaning solution bag 125, open the thimble valves B8, B6, B5, and B12, pass air from the fifth air channel A5, and push the secondary cleaning solution into the magnetic bead capture chamber 108 first, The magnetic beads with biomacromolecular extracts such as nucleic acid that are uniformly adsorbed on the inner wall surface of the magnetic bead capture chamber 108 are fully washed and then pushed into the waste liquid chamber 107 . Then, air is continuously introduced from the fifth air channel A5 to dry up the reagent remaining on the surface of the magnetic beads in the magnetic bead capture chamber 108, and then the thimble valves B8, B6, B5, and B12 are closed. In this step, if it is necessary to measure a certain amount of cleaning solution, it can be realized through the second quantitative cavity 106, and its specific operation is the same as that of measuring a certain amount of lysate, and will not be repeated here.

The eighth step, release the eluent: puncture the eluent bag 124, open the thimble valves B2, B4, B13, B14, pass air from the fourth flow channel A4 to push the eluent into the mixing chamber 109, and continue Air is continuously introduced from the fourth air channel A4, and the eluent is mixed with the freeze-dried beads in the mixing chamber 109 by the air bubbles generated from the bottom of the mixing chamber 109 to form an elution mixture, and then the thimble valves B13 and B14 are closed .

Step 9: Take a certain amount of eluent mixture and push it into the PCR chamber: open the thimble valve B15, and let air flow through the eighth airway A8 to push a part of the eluate mixture into the first quantification chamber 110. Use the detection module of the supporting instrument to judge that the eluent mixture with a predetermined volume (25 μL) has been measured in the first quantitative chamber 110, and then close the thimble valves B2, B4, and B15. Open the thimble valve B16, pass air from the ninth airway A9 to push the eluate mixture collected in the first quantitative chamber 110 into the No. 1 PCR chamber 111, and then close the thimble valve B16.

Re-open the thimble valves B2, B4, and B15, and pass air from the eighth airway A8 to push a part of the eluate mixture into the first quantitative chamber 110. Use the detection module of the supporting instrument to judge that the eluent mixture with a predetermined volume (25 μL) has been measured in the first quantitative chamber 110, and then close the thimble valves B2, B4, and B15. Open the thimble valve B17, pass air from the ninth airway A9 to push the eluate mixture collected in the first quantitative chamber 110 into the No. 2 PCR chamber 112, and then close the thimble valve B17.

Re-open the thimble valves B2, B4, and B15, and pass air from the eighth airway A8 to push a part of the eluate mixture into the first quantitative chamber 110. Use the detection module of the supporting instrument to judge that the eluent mixture with a predetermined volume (25 μL) has been measured in the first quantitative chamber 110, and then close the thimble valves B2, B4, and B15. Open the thimble valve B18, pass air from the ninth airway A9 to push the eluate mixture collected in the first quantitative chamber 110 into the No. 3 PCR chamber 113, and then close the thimble valve B18.

Re-open the thimble valves B2, B4, and B15, and pass air from the eighth airway A8 to push a part of the eluate mixture into the first quantitative chamber 110. Use the detection module of the supporting instrument to judge that the eluent mixture with a predetermined volume (25 μL) has been measured in the first quantitative chamber 110, and then close the thimble valves B2, B4, and B15. Open the thimble valve B19, pass air from the ninth airway A9 to push the eluate mixture collected in the first quantitative chamber 110 into the No. 4 PCR chamber 114, and then close the thimble valve B19.

Step 10: Use the amplification heating module of the supporting instrument to perform a temperature rise and fall cycle on the 4 PCR chambers, and detect the amplification result through the amplification detection module. After the experiment is over, the top area (the tenth airway A10 near its vent) and the bottom area (the sixth flowway 146 is located in the second quantitative chamber 110 and the thimble valve B16) of the four PCR chambers are hot-melt plastic-sealed by supporting instruments. The channel section between), so as to seal the PCR cavity, prevent the amplification product from leaking out of the cartridge along the flow channel, and pollute the supporting instruments and the environment.

To sum up, the present invention proposes a microfluidic nucleic acid extraction, amplification and detection integrated cartridge and detection device. The reaction chambers such as the sample chamber, the magnetic bead storage chamber 105, the magnetic bead capture chamber 108, the waste liquid chamber 107, the mixing The uniform chamber 109 and the PCR chamber are reasonably arranged on the cartridge body 1, and the reagents required for the reaction are independently sealed in the reagent pack also arranged on the cartridge body 1, which solves the problem of reagent storage. The piercing of the reagent pack is preferably achieved by using a piercing needle structure.

In addition, the cartridge body 1 is reasonably designed with an air flow channel, a main channel and a valve (thimble valve). The flow direction and stop position of the sample and reagent are controlled by the thimble valve switch, and the flow direction and stop position of the sample and reagent are controlled by the way of driving the liquid by the air pump to control the inside of the cartridge body 1. The flow of the liquid enables the reagent package to be selectively communicated with the reaction chamber, and the control is simple and reliable.

Moreover, when the liquid in the cartridge is controlled by an air pump, or when the liquid reacts in the reaction chamber, air bubbles are easily generated. The prior art adopts a cartridge with a horizontal structure, and the above-mentioned reaction or the air bubbles generated when the air pump is propelled are not easy to remove. However, the cartridge body 1 of the present invention has a vertical structure, and the bubbles generated inside will rise from the bottom to the top during the reaction, which can be easily eliminated.

Therefore, a microfluidic nucleic acid extraction, amplification and detection integrated cartridge and device proposed in the present invention can realize automatic processing of nucleic acid extraction, amplification, and detection, and the detection results are more accurate, and only a few manual steps are required. Simple operation, safe and convenient. Using the device for detection, the detection time is less than one hour, and the manual operation does not exceed 2 minutes, and it can be used to detect pathogen genome targets of various human clinical samples, and has wide applicability. The storage and transportation of the device of this application are all under normal temperature conditions, no cold chain is needed, and the economy is high. The cartridge of the device of the present invention is completely closed during the detection process, and its detection result is completely consistent with the conventional method, and can be operated without professional training, which is safe and convenient.

The above is only a preferred embodiment of the present invention, it should be pointed out that for those of ordinary skill in the art, without departing from the technical principle of the present invention, some improvements and replacements can also be made, these improvements and replacements It should also be regarded as the protection scope of the present invention.

Claims (15)

1. The card box is characterized by comprising a card box body (1), a reagent pack group (2), a sealing cover (3) and a waterproof breathable film (4);

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 preset positions 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, the other end of the air flow channel is provided with an air vent for connecting an air pump, and the waterproof air-permeable membrane (4) is used for sealing the air vent;

the reaction cavity comprises a sample cavity, a magnetic bead storage cavity (105), a magnetic bead capture cavity (108), a waste liquid cavity (107), a mixing cavity (109), a first quantitative cavity (110) and a PCR cavity; the sample cavity is respectively communicated with the magnetic bead storage cavity (105) and the first end of the magnetic bead capture cavity (108); the first end of the magnetic bead capturing cavity (108) is also communicated with the uniform mixing cavity (109), and the opposite second end of the magnetic bead capturing cavity is communicated with the waste liquid cavity (107); the mixing cavity (109) is communicated with a first end of the first quantitative cavity (110), and an opposite second end of the first quantitative cavity (110) is communicated with the PCR cavity; the sample cavity is provided with a sample adding opening (5), and the cover (3) is used for sealing the sample adding opening (5);

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 four reagent pack units are respectively a lysis solution pack (121), a binding solution pack (122), a first cleaning solution pack (123) and an eluent pack (124);

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 the first gas flow channel (A1) corresponds to the lysis liquid pack (121) and the second gas flow channel (A2) corresponds to the binding liquid pack (122); the main flow passage comprises a first branch flow passage (141) and a second branch flow passage (142);

two ends of the first branch flow channel (141) are respectively communicated with the bottom end of the sample cavity and the bottom end of the first air flow channel (A1);

one end of the second branch flow channel (142) is communicated with the bottom end of the sample cavity, the other end of the second branch flow channel is connected to the first branch flow channel (141), the connection position is a first connection point (C1), and two ends of the magnetic bead storage cavity (105) are respectively connected to the second branch flow channel (142);

the bottom end of the second air flow channel (A2) is connected to a pipe section of the first branch flow channel (141) between the first air flow channel (A1) and the first connection point (C1);

the valves are arranged at preset positions of the first branch flow channel (141) and the second branch flow channel (142) to control the lysis solution in the lysis solution bag (121) and the binding solution in the binding solution bag (122) to directly flow into the sample cavity or sequentially flow through the magnetic bead storage cavity (105) and the sample cavity.

3. A cartridge according to claim 2, wherein the set of reagent packs (2) further comprises a second wash pack (125) and a reserve pack (126); the air flow channel also comprises a fifth air flow channel (A5) and a sixth air flow channel (A6);

the third air flow channel (A3) corresponds to the first cleaning solution bag (123), and the bottom end of the third air flow channel (A3) is connected to a pipe section, which is positioned between the second air flow channel (A2) and the first connecting point (C1), on the first branch flow channel (141);

the fifth air flow channel (A5) corresponds to the second cleaning solution bag (125), and the bottom end of the fifth air flow channel (A5) is connected to a pipe section, which is positioned between the third air flow channel (A3) and the first connecting point (C1), on the first branch flow channel (141);

the sixth gas flow channel (A6) corresponds to the reserved liquid bag (126), and the bottom end of the sixth gas flow channel (A6) is connected to a pipe section, which is positioned between the fifth gas flow channel (A5) and the first connecting point (C1), on the first branch flow channel (141).

4. A cartridge according to claim 3, wherein the reaction chamber further comprises a second dosing chamber (106); a seventh air passage (A7) is arranged at the top of the second quantitative cavity (106), and the other end of the seventh air passage (A7) is provided with the air vent; the bottom of the second quantitative cavity (106) is connected to a pipe section, which is positioned between the sixth gas flow channel (A6) and the first connecting point (C1), of the first branch flow channel (141), and the two sides of the connecting position are respectively provided with the valves.

5. The cartridge of claim 2, wherein the main flow channel further comprises a third branch flow channel (143), a predetermined area section of the third branch flow channel (143) forming the magnetic bead capture chamber (108); a first end of the third branch flow channel (143) is connected to the first connecting point (C1), a second end opposite to the first end is communicated with one end of the waste liquid cavity (107), and the other end of the waste liquid cavity (107) is provided with an air outlet channel;

the fourth gas flow channel (A4) corresponds to the eluent bag (124), and the bottom end of the fourth gas flow channel (A4) is connected to the second end of the third branch flow channel (143);

the valve is arranged at a preset position of the third branch flow channel (143) to control the liquid in the magnetic bead capture cavity (108) to flow into the waste liquid cavity (107) from the second end of the third branch flow channel (143), and control the eluent in the eluent bag (124) to flow into the magnetic bead capture cavity (108) from the second end of the third branch flow channel (143).

6. The cartridge of claim 2, wherein the sample chamber comprises a sample addition chamber (101) and a sample processing chamber (103), the sample addition chamber (101) and the sample processing chamber (103) being in communication via a siphon trap (102); one end of the siphon elbow (102) is communicated with the bottom of the sample adding cavity (101), and the other end of the siphon elbow is communicated with the top of the sample processing cavity (103); the top of the sample adding cavity (101) is provided with the sample adding port (5); an eighth air channel (A8) is arranged at the top of the sample processing cavity (103).

7. A cartridge according to claim 6, wherein a condensation chamber (104) is provided at a predetermined position of the eighth air passage (A8), the condensation chamber (104) having a water-absorbing agent pre-disposed therein.

8. The cartridge of claim 6, wherein the main flow channel further comprises a fourth branch flow channel (144), a fifth branch flow channel (145) and a sixth branch flow channel (146);

one end of the fourth branch flow channel (144) is communicated with the bottom of the sample processing cavity (103), and the other end of the fourth branch flow channel is communicated with the top of the uniform mixing cavity (109);

one end of the fifth branch flow channel (145) is communicated with the first end of the third branch flow channel (143), and the other end of the fifth branch flow channel is communicated with the first end of the uniform mixing cavity (109);

one end of the sixth branch flow channel (146) is communicated with the opposite second end of the blending cavity (109), and the other end of the sixth branch flow channel is communicated with the PCR cavity; a tenth air passage (A10) is arranged at the top of the PCR cavity;

a ninth air passage (A9) is arranged at the top of the first quantitative cavity (110), and the other end of the ninth air passage (A9) is provided with the air vent; the bottom of the first quantitative cavity (110) is connected to a pipe section, which is positioned between the blending cavity (109) and the PCR cavity, of the sixth branch flow channel (146), and the two sides of the connection position are respectively provided with the valves.

9. The cartridge of claim 8, wherein the PCR chambers are provided in number side by side, and the bottom of each PCR chamber is provided with the valve so as to be selectively communicated with the first quantitative chamber (110).

10. A cartridge according to any of claims 1-9, wherein the valve is a spike valve (6); the inside of needle ejection valve (6) has the passageway, be equipped with seal membrane (61) on the passageway, take place to warp when seal membrane (61) receive predetermined pressure to seal the passageway.

11. A cartridge according to claim 1, wherein the reagent pack set (2) comprises a housing (21) and a sealing membrane (22); at least four liquid containing cavities (23) are arranged on the shell (21), corresponding test solutions are preset in the liquid containing cavities (23), and a circle of sticking grooves (24) are formed in the edges of the liquid containing cavities (23); the number of the sealing films (22) corresponds to the number of the liquid containing cavities (23), and the sealing films (22) are attached to the attaching grooves (24) to seal the liquid containing cavities (23), so that the reagent pack unit is formed.

12. A cartridge according to claim 11, wherein the flow channel comprises an upper flow channel (131), a test solution chamber (132) and a lower flow channel (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 accommodating cavity (23), and the reagent pack unit is broken and then the built-in test solution flows into the test solution cavity (132) and the lower runner (133) in sequence.

13. A cartridge according to claim 12, wherein a lancet (9) is provided in the test solution chamber (132) at a predetermined distance from the sealing membrane (22).

14. 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 plate (11) is provided with a plurality of concave cavities, and the front film (12) is attached to the front surface of the cassette plate (11) to seal the concave cavities, so that other reaction cavities except the PCR cavity are formed;

the cassette board (11) is provided with a plurality of through holes, 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 through holes, so that the PCR cavity is formed;

the cassette board (11) is provided with a plurality of grooves, and the front film (12) is attached to the front surface of the cassette board (11) to seal the grooves, so that the airflow channel and the main flow channel are formed.

15. A testing device comprising a cartridge according to any one of claims 1 to 14 and associated apparatus.

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