CN115369023A

CN115369023A - Magnetic bead cleaning structure and cleaning method

Info

Publication number: CN115369023A
Application number: CN202110564085.6A
Authority: CN
Inventors: 梅哲; 贾础豪; 张彤
Original assignee: Guangzhou Wondfo Biotech Co Ltd
Current assignee: Guangzhou Wondfo Biotech Co Ltd
Priority date: 2021-05-21
Filing date: 2021-05-21
Publication date: 2022-11-22

Abstract

The invention relates to the technical field of biomedical treatment, and discloses a magnetic bead cleaning structure and a cleaning method, wherein the magnetic bead cleaning structure comprises a card box body, and a sample cavity, a magnetic bead capturing cavity, a waste liquid cavity, a sixth air passage, an eighth air passage, a first cleaning liquid bag and a third air passage which are arranged on the card box body; the sample cavity is selectively communicated with a first end of the magnetic bead capture cavity, and an opposite second end of the magnetic bead capture cavity is selectively communicated with the waste liquid cavity; and one ends of the sixth air passage, the eighth air passage and the third air passage are provided with air vents, the other end of the sixth air passage is communicated with the top of the sample cavity, the other end of the eighth air passage is communicated with the upper part of the waste liquid cavity, the other end of the third air passage is selectively communicated with the bottom of the sample cavity, the third air passage corresponds to the first cleaning liquid bag, and cleaning liquid in the first cleaning liquid bag is sequentially flowed into the third air passage and the sample cavity after the first cleaning liquid bag is broken. The invention can ensure that the magnetic beads are cleaned more thoroughly, and can blow the cleaning liquid remained in the cavity and on the magnetic beads to dry.

Description

Magnetic bead cleaning structure and cleaning method

Technical Field

The invention relates to the technical field of biological medical treatment, in particular to a magnetic bead cleaning structure and a cleaning method.

Background

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

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

In the prior art, due to the limitation of a cavity or a flow channel structure, magnetic beads are not cleaned thoroughly easily, and residual cleaning liquid in the cavity and on the magnetic beads is not dried after the magnetic beads are cleaned, so that subsequent eluent is mixed with the residual cleaning liquid, and the elution effect is influenced.

Thus, improvements in the prior art are needed.

Disclosure of Invention

The purpose of the invention is: the invention provides a magnetic bead cleaning structure and a cleaning method, and aims to solve the technical problems that in the prior art, due to the limitation of a cavity or a flow channel structure, magnetic beads are easy to clean incompletely, and after the magnetic beads are cleaned, residual cleaning liquid in the cavity and on the magnetic beads is not dried, so that subsequent eluent is mixed with the residual cleaning liquid, and the elution effect is influenced.

In order to achieve the purpose, the invention provides a magnetic bead cleaning structure, which comprises a card box body, a sample cavity, a magnetic bead capturing cavity, a waste liquid cavity, a sixth air channel, an eighth air channel, a first cleaning liquid bag and a third air channel, wherein the sample cavity, the magnetic bead capturing cavity, the waste liquid cavity, the sixth air channel, the eighth air channel, the first cleaning liquid bag and the third air channel are arranged on the card box body;

the sample cavity is selectively communicated with a first end of the magnetic bead capture cavity, and an opposite second end of the magnetic bead capture cavity is selectively communicated with the waste liquid cavity;

and one ends of the sixth air passage, the eighth air passage and the third air passage are provided with air vents, the other end of the sixth air passage is communicated with the top of the sample cavity, the other end of the eighth air passage is communicated with the upper part of the waste liquid cavity, the other end of the third air passage is selectively communicated with the bottom of the sample cavity, the third air passage corresponds to the first cleaning liquid bag, and cleaning liquid arranged in the first cleaning liquid bag after the first cleaning liquid bag is broken sequentially flows into the third air passage and the sample cavity.

In some embodiments of the present application, the cleaning device further comprises a second cleaning solution bag and a fifth air flow channel corresponding to the second cleaning solution bag; one end of the fifth air flow channel is provided with an air vent, the other end of the fifth air flow channel is selectively communicated with the bottom of the sample cavity, and secondary cleaning liquid which is arranged in the fifth air flow channel (A5) and the sample cavity flows into the fifth air flow channel (A5) and the sample cavity in sequence after the second cleaning liquid is wrapped and broken.

In some embodiments of the present application, the apparatus further comprises a mixing chamber, a PCR chamber, and a seventh airway; one end of the mixing cavity is communicated with the first end of the magnetic bead capturing cavity, and the other end of the mixing cavity is communicated with the PCR cavity; and one end of the seventh air passage is provided with an air vent, and the other end of the seventh air passage is communicated with the top of the uniform mixing cavity.

In some embodiments of the present application, the sample chamber includes a sample addition chamber and a sample processing chamber, and the sample addition chamber is communicated with the sample processing chamber through a siphon elbow; one end of the siphon bent pipe is communicated with the bottom of the sample adding cavity, and the other end of the siphon bent pipe is communicated with the top of the sample processing cavity; the sixth air channel is communicated with the top of the sample processing cavity.

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

In some embodiments of the present application, the third flow channel includes an upper air channel, a test solution cavity, and a lower flow channel; one end of the upper air passage is provided with the vent, and the other end of the upper air passage is communicated with the upper part of the test solution cavity; one end of the lower flow channel is communicated with the lower part of the test solution cavity, and the other end of the lower flow channel is connected into the sample cavity; and after the first cleaning liquid bag is broken, the built-in test liquid sequentially flows into the test liquid cavity and the lower runner.

The application also provides a magnetic bead cleaning method based on the magnetic bead cleaning structure of any one of the preceding claims, comprising the following steps:

s1, air vents of the sixth air passage, the eighth air passage and the third air passage are all connected with an air pump of a matched instrument; a magnetic attracting module is started by a matched instrument at a position corresponding to the sample cavity, and magnetic beads are attracted on the inner wall of the sample cavity;

s2, introducing air from the sixth air channel, and sequentially pushing the mixed solution in the sample cavity into the magnetic bead capturing cavity and the waste liquid cavity;

s3, puncturing the first cleaning liquid bag, and enabling cleaning liquid to flow out of the first cleaning liquid bag to the third airflow channel;

s4, introducing air from the third airflow channel, and pushing a cleaning solution into the sample cavity;

and S5, the magnetic attraction module is closed by the matched instrument, and air is continuously introduced from the third air flow channel.

In some embodiments of the present application, the method further comprises the steps of:

s6, starting a magnetic attraction module at a position corresponding to the magnetic bead capture cavity by a matched instrument;

s7, introducing air from the sixth air channel, and sequentially pushing the mixed solution in the sample cavity into the magnetic bead capturing cavity and the waste liquid cavity; the magnetic attraction module adsorbs magnetic beads in liquid to the inner wall of the magnetic bead capture cavity;

s8, puncturing the second cleaning liquid bag to enable secondary cleaning liquid to flow out of the second cleaning liquid bag to the fifth airflow channel;

s9, introducing air from the fifth airflow channel, and pushing a secondary cleaning solution into the sample cavity;

s10, introducing air from the sixth air channel, and sequentially pushing the secondary cleaning liquid in the sample cavity into the magnetic bead capturing cavity and the waste liquid cavity;

s11, continuously introducing air from the sixth air channel to blow dry the magnetic beads in the magnetic bead capturing cavity.

The application also provides another magnetic bead cleaning method based on the magnetic bead cleaning structure of any one of the preceding claims, which comprises the following steps:

s101, air vents of the sixth air channel, the eighth air channel and the third air channel are all connected with an air pump of a matched instrument; the magnetic attraction module is started by a matched instrument at a position corresponding to the magnetic bead capture cavity;

s102, introducing air from the sixth air channel, and pushing all the mixed liquid in the sample cavity into the waste liquid cavity; the magnetic attraction module adsorbs magnetic beads in the mixed liquid to the inner wall of the magnetic bead capture cavity;

s103, puncturing the first cleaning liquid bag, and enabling cleaning liquid to flow out of the first cleaning liquid bag to the third airflow channel;

s104, introducing air from the third airflow channel, and pushing a cleaning solution into the sample cavity;

s105, introducing air from the sixth air channel, and sequentially pushing the cleaning liquid in the sample cavity into the magnetic bead capturing cavity and the waste liquid cavity.

s106, puncturing the second cleaning liquid bag, and enabling secondary cleaning liquid to flow out of the second cleaning liquid bag to the fifth airflow channel;

s107, introducing air from the fifth airflow channel, and pushing a secondary cleaning solution into the sample cavity;

s108, introducing air from the sixth air channel, and sequentially pushing the secondary cleaning liquid in the sample cavity into the magnetic bead capturing cavity and the waste liquid cavity;

and S109, continuously introducing air from the sixth air channel to blow dry the magnetic beads in the magnetic bead capturing cavity.

Compared with the prior art, the magnetic bead cleaning structure and the cleaning method provided by the embodiment of the invention have the beneficial effects that:

according to the magnetic bead cleaning structure and the cleaning method provided by the embodiment of the invention, the structure and the connection mode of the chamber and the ventilation flow channel are improved, so that the magnetic beads are cleaned more thoroughly, air can be continuously introduced through the ventilation flow channel after the magnetic beads are cleaned, the cleaning liquid remained in the chamber and on the magnetic beads is dried, and the subsequent eluent is prevented from being mixed with the remained cleaning liquid to influence the elution effect.

Drawings

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

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

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

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

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

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

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

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

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

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

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

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

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

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

FIG. 14 is a schematic structural view of the gas flow passage region of FIG. 4;

in the figure:

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

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

2. a reagent pack group; 21. a housing; 22. sealing the film; 23. attaching an accessory; 24. a limiting strip; 121. a lysis solution bag; 122. combining the liquid bag; 123. a first cleaning solution bag; 124 an eluate package; 125. a second cleaning liquid bag;

3. sealing the cover; 4. breaking the package;

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

131. an upper airway; 132. a test solution chamber; 133. a lower runner;

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

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

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

Detailed Description

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

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

It is to be understood that the terms "length," "width," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in an orientation or positional relationship that is indicated based on the orientation or positional relationship as shown in the figures, which is for convenience in describing the invention and to simplify the description, and that does not indicate or imply that the device or element so referred to must have a particular orientation, be constructed in a particular orientation, and be operated in a particular manner and is not to be construed as limiting the invention.

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

As shown in fig. 1 to 4, a cartridge according to a preferred embodiment of the present invention includes a cartridge body 1, a reagent pack group 2, and a cover 3.

Specifically, the card box body 1 is provided with a main flow channel, a valve group, a reaction cavity and an airflow channel. The valve set includes several valves set in the preset position of the main flow passage to make the reaction cavity and one end of the airflow passage communicated selectively with the main flow passage and the other end of the airflow passage has air port for connecting the air pump.

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

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

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

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

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

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

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

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

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

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

In the above steps, the valve set and the air pump control the flow of the liquid.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

In the above embodiment, referring to FIGS. 10-12, PCR chamber 109 comprises central chamber 161, amplification chamber 162, zone flow channel 163, central flow channel groove 164, gas containment zone 165, waterproof gas permeable membrane 166, zone gas channel 167, gas distribution channel 168, and transition gas zone 169.

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

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

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

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

In the above embodiment, referring to fig. 1, the reagent pack group 2 is attached to the back surface of the left upper side of the cassette board 11, and the reagent pack group 2 includes the housing 21, the sealing film 22, and the attachment 23. The casing 21 is provided with five liquid containing cavities with openings facing the front, and the sealing film 22 is attached to the front of the casing 21 to seal the liquid containing cavities, so as to form five reagent pack units, namely a corresponding lysis solution pack 121, a binding solution pack 122, a first cleaning solution pack 123, an elution solution pack 124 and a second cleaning solution pack 125. One surface of the attachment 23 is used for connecting the front surface of the shell 21, the other surface of the attachment 23 is used for connecting the back surface of the cassette board 11, and a hollow part is formed at a position corresponding to the liquid containing cavity on the attachment 23, namely, the attachment 23 does not shield the sealing film 22.

In some embodiments of the present application, a bale breaker 4 is also included. The bag breaking piece 4 is arranged at a position with a preset distance from the sealing film 22, and the bag breaking piece 4 breaks the sealing film 22 when being subjected to preset pressure.

In some embodiments of the present application, referring to fig. 3 and 7, the back of the cassette board 11 is provided with a plurality of position-limiting strips 24 conforming to the shape of the housing 21, and the edges of the attachment 23 are located in the position-limiting strips 24 to prevent the reagent pack set 2 from shifting.

In some embodiments of the present application, referring to fig. 14, each of the first air flow channel A1, the second air flow channel A2, the third air flow channel A3, the fourth air flow channel A4, and the fifth air flow channel A5 includes an upper air channel 131, a test solution cavity 132, and a lower air channel 133, one end of the upper air channel 131 has an air vent, and the other end thereof communicates with an upper portion of the test solution cavity 132. One end of the lower channel 133 communicates with the lower portion of the sample chamber 132, and the other end thereof is connected to the main channel. The test solution chamber 132 corresponds to the solution containing chamber, and the reagent pack unit is broken and the built-in test solution flows into the test solution chamber 132 and the lower flow channel 133 in sequence.

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

The working process of the cartridge structure provided by the above embodiment is as follows:

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

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

Step three, sample pretreatment: sample pretreatment: after sample application, the sample is mixed with the internal reference lyophilized beads in the internal reference lyophilized bead cavity, and then reaches the siphon elbow 110 to stay. After the start of the test, the sample staying in the siphon tube 110 is drawn into the sample processing chamber 103 by using the kit pneumatic pump to draw air from the air vent of the sixth air passage A6 (the following description will not refer to the operation of the kit pneumatic pump, and only the air vent or air suction state of the air passage will be described).

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

Step five, releasing the binding solution: the binding liquid pack 122 is punctured, and the binding liquid flows out from the binding liquid pack 122 to the second gas flow channel A2. The second valve B2 is opened and air is introduced from the second flow channel A2 to push the binding solution into the sample processing chamber 103. And continuously introducing air from the second air flow channel A2, mixing the binding solution and the magnetic beads with the lysed sample by using the bubbles continuously generated from the bottom of the sample processing chamber 103, and then closing the second valve B2.

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

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

The first magnetic bead cleaning mode is to adsorb the magnetic bead in the magnetic bead capturing cavity 106 for cleaning, and the magnetic module is only arranged at the corresponding position of the magnetic bead capturing cavity 106 by the matched instrument. The method comprises the following specific steps: and opening the third valve B3 and the sixth valve B6, and simultaneously starting the magnetic suction module by a matched instrument. Air is introduced from the sixth air channel A6, and the mixed solution in the sample processing chamber 103 is sequentially pushed into the magnetic bead capture chamber 106 and the waste liquid chamber 105. When the mixed solution passes through the magnetic bead capturing cavity 106, the magnetic beads are uniformly adsorbed on the inner wall of the magnetic bead capturing cavity 106 under the action of the magnetic attraction force of the magnetic attraction module, and the rest liquid part of the mixed solution is completely pushed into the waste liquid cavity 105, and then the third valve B3 and the sixth valve B6 are closed. The first cleaning liquid pack 123 is punctured, so that the cleaning liquid flows out from the first cleaning liquid pack 123 to the third air flow channel A3. The fourth valve B4 and the fifth valve B5 are opened, air is introduced from the third air flow channel A3, the wash liquid is pushed into the sample processing chamber 103, and then the fourth valve B4 and the fifth valve B5 are closed. And opening a third valve B3 and a sixth valve B6, introducing air from a sixth air channel A6, and sequentially pushing the cleaning solution into the magnetic bead capturing cavity 106 and the waste liquid cavity 105 to finish the first magnetic bead cleaning. The second cleaning solution bag 125 is continuously punctured, the fifth valve B5 is opened, air is introduced from the fifth air flow channel A5, the cleaning solution is pushed into the sample processing chamber 103, and then the fifth valve B5 is closed. And opening a third valve B3 and a sixth valve B6, introducing air from a sixth air channel A6, and sequentially pushing the cleaning solution into the magnetic bead capturing cavity 106 and the waste liquid cavity 105 to finish the second magnetic bead cleaning. And continuously introducing air from the sixth air channel A6 to blow dry the magnetic bead capture cavity 106.

The second kind of magnetic bead cleaning method is to adsorb the magnetic bead and carry out a washing in sample processing chamber 103, carries out the secondary and washs in magnetic bead capture chamber 106, and supporting instrument all sets up magnetism and inhales the module in magnetic bead capture chamber 106 and sample processing chamber 103 corresponding position department. The method comprises the following specific steps: the matched instrument starts the magnetic module at the corresponding position of the sample processing cavity 103, opens the third valve B3 and the sixth valve B6, and introduces air from the sixth air passage A6. Under the magnetic attraction of the magnetic module, the magnetic beads are uniformly adsorbed on the inner wall of the sample processing chamber 103, and the rest of the liquid of the mixed liquid is completely pushed into the waste liquid chamber 105, and then the third valve B3 and the sixth valve B6 are closed. The first cleaning liquid pack 123 is punctured, so that the cleaning liquid flows out from the first cleaning liquid pack 123 to the third air flow channel A3. The fourth valve B4 and the fifth valve B5 are opened, and air is introduced from the third air flow channel A3 to push the wash solution into the sample processing chamber 103. Then the magnetic module is closed by the matching instrument, air is continuously introduced from the third air flow channel A3, bubbles continuously generated from the bottom of the sample processing chamber 103 are utilized to fully mix the cleaning solution and the magnetic beads, and then the fourth valve B4 and the fifth valve B5 are closed. The magnetic module located at the corresponding position of the magnetic bead capture cavity 106 is opened by the matching instrument, and the third valve B3 and the sixth valve B6 are opened to introduce air from the sixth air channel A6. Under the magnetic attraction of the magnetic module, the magnetic beads are uniformly adsorbed on the inner wall of the magnetic bead capturing cavity 106, the rest liquid part of the mixed liquid is completely pushed into the waste liquid cavity 105, and then the third valve B3 and the sixth valve B6 are closed. The second cleaning solution bag 125 is continuously punctured, the fifth valve B5 is opened, air is introduced from the fifth air flow channel A5, the cleaning solution is pushed into the sample processing chamber 103, and then the fifth valve B5 is closed. And opening a third valve B3 and a sixth valve B6, introducing air from a sixth air channel A6, and sequentially pushing the cleaning solution into the magnetic bead capturing cavity 106 and the waste liquid cavity 105 to finish the second magnetic bead cleaning. And continuously introducing air from the sixth air channel A6 to blow dry the reagent remained on the surface of the magnetic bead in the magnetic bead capturing cavity 106.

Seventh step, releasing the eluent: the eluent packet 124 is punctured such that eluent flows out of the eluent packet 124 to the fourth gas flow channel A4. And opening a seventh valve B7 and an eighth valve B8, introducing air from the fourth air flow channel A4, and sequentially pushing the eluent into the magnetic bead capturing cavity 106 and the mixing cavity 107. And continuously introducing air from the fourth air flow channel A4, mixing the eluent with the freeze-dried beads in the blending cavity 107 by utilizing the air bubbles continuously generated from the bottom of the blending cavity 107, and then closing the seventh valve B7 and the eighth valve B8.

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

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

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

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

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

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

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

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

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

Claims

1. A magnetic bead cleaning structure is characterized by comprising a card box body (1), and a sample cavity, a magnetic bead capturing cavity (106), a waste liquid cavity (105), a sixth air channel (A6), an eighth air channel (A8), a first cleaning liquid bag (123) and a third air channel (A3) which are arranged on the card box body (1);

the sample chamber is selectively communicated with a first end of the magnetic bead capture chamber (106), and an opposite second end of the magnetic bead capture chamber (106) is selectively communicated with the waste liquid chamber (105);

one end of the sixth air passage (A6), one end of the eighth air passage (A8) and one end of the third air passage (A3) are provided with air vents, the other end of the sixth air passage (A6) is communicated with the top of the sample cavity, the other end of the eighth air passage (A8) is communicated with the upper part of the waste liquid cavity (105), the other end of the third air passage (A3) is selectively communicated with the bottom of the sample cavity, the third air passage (A3) corresponds to the first cleaning liquid bag (123), and cleaning liquid contained in the first cleaning liquid bag (123) flows into the third air passage (A3) and the sample cavity in sequence after the bag is broken.

2. The magnetic bead washing structure of claim 1, further comprising a second washing solution pack (125) and a fifth gas flow channel (A5) corresponding to the second washing solution pack (125); one end of the fifth air flow channel (A5) is provided with an air vent, the other end of the fifth air flow channel is selectively communicated with the bottom of the sample cavity, and secondary cleaning liquid arranged in the second cleaning liquid bag (125) flows into the fifth air flow channel (A5) and the sample cavity in sequence after the second cleaning liquid bag is broken.

3. The magnetic bead washing structure of claim 2, further comprising a mixing chamber (107), a PCR chamber (109), and a seventh air channel (A7); one end of the mixing cavity (107) is communicated with the first end of the magnetic bead capture cavity (106), and the other end of the mixing cavity is communicated with the PCR cavity (109); one end of the seventh air passage (A7) is provided with an air vent, and the other end of the seventh air passage is communicated with the top of the blending cavity (107).

4. The magnetic bead washing structure of claim 2, wherein the sample chamber comprises a sample addition chamber (101) and a sample processing chamber (103), and the sample addition chamber (101) is communicated with the sample processing chamber (103) through a siphon elbow (110); one end of the siphon elbow (110) 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 sixth air passage (A6) is communicated with the top of the sample processing cavity (103).

5. The magnetic bead washing structure of claim 4, characterized in that an internal reference freeze-dried bead chamber (102) is further included between the sample addition chamber (101) and the sample processing chamber (103); one end of the internal reference freeze-dried bead cavity (102) is communicated with the bottom of the sample adding cavity (101), and the other end of the internal reference freeze-dried bead cavity is communicated with one end of the siphon elbow (111).

6. The magnetic bead washing structure of claim 2, wherein the third flow channel (A3) comprises an upper flow channel (131), a sample 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 sample cavity; after the first cleaning liquid bag (123) is broken, the built-in test liquid flows into the test liquid cavity (132) and the lower runner (133) in sequence.

7. A method for washing magnetic beads, which is based on the magnetic bead washing structure as claimed in any one of claims 2-6, and comprises the following steps:

s1, air vents of the sixth air channel (A6), the eighth air channel (A8) and the third air channel (A3) are all connected with an air pump of a matched instrument; the matched instrument starts the magnetic attraction module at the corresponding position of the sample cavity to attract the magnetic beads on the inner wall of the sample cavity;

s2, introducing air from the sixth air channel (A6), and sequentially pushing the mixed solution in the sample cavity into the magnetic bead capture cavity (106) and the waste liquid cavity (105);

s3, puncturing the first cleaning liquid bag (123) to enable cleaning liquid to flow out of the first cleaning liquid bag (123) to the third air flow channel (A3);

s4, introducing air from the third air flow channel (A3) and pushing a cleaning solution into the sample cavity;

s5, the magnetic suction module is closed by the matched instrument, and air is continuously introduced from the third air flow channel (A3).

8. The method of claim 7, further comprising the steps of:

s6, starting a magnetic attraction module at a position corresponding to the magnetic bead capture cavity (106) by a matched instrument;

s7, introducing air from the sixth air channel (A6), and sequentially pushing the mixed solution in the sample cavity into the magnetic bead capture cavity (106) and the waste liquid cavity (105); the magnetic attraction module adsorbs magnetic beads in liquid to the inner wall of the magnetic bead capture cavity (106);

s8, puncturing the second cleaning liquid bag (125) to enable the secondary cleaning liquid to flow out of the second cleaning liquid bag (125) to the fifth air flow channel (A5);

s9, introducing air from the fifth airflow channel (A5), and pushing a secondary cleaning solution into the sample cavity;

s10, introducing air from the sixth air channel (A6), and sequentially pushing the secondary cleaning solution in the sample cavity into the magnetic bead capture cavity (106) and the waste liquid cavity (105);

s11, continuously introducing air from the sixth air channel (A6) to blow dry the magnetic beads in the magnetic bead capturing cavity (106).

9. A method for washing magnetic beads, which is based on the magnetic bead washing structure as set forth in any of claims 2-6, and comprises the following steps:

s101, air vents of the sixth air channel (A6), the eighth air channel (A8) and the third air channel (A3) are all connected with an air pump of a matched instrument; a magnetic attracting module is started by a matched instrument at a position corresponding to the magnetic bead capturing cavity (106);

s102, introducing air from the sixth air channel (A6) and pushing all the mixed solution in the sample cavity into the waste liquid cavity (105); the magnetic attraction module adsorbs magnetic beads in the mixed solution to the inner wall of the magnetic bead capture cavity (106);

s103, puncturing the first cleaning liquid bag (123) to enable cleaning liquid to flow out of the first cleaning liquid bag (123) to the third air flow channel (A3);

s104, introducing air from the third air flow channel (A3) and pushing a cleaning solution into the sample cavity;

s105, introducing air from the sixth air channel (A6), and sequentially pushing the cleaning solution in the sample cavity into the magnetic bead capture cavity (106) and the waste liquid cavity (105).

10. A method for washing a magnetic bead as claimed in claim 9, further comprising the steps of:

s106, puncturing the second cleaning liquid bag (125) to enable the secondary cleaning liquid to flow out of the second cleaning liquid bag (125) to the fifth airflow channel (A5);

s107, introducing air from the fifth airflow channel (A5), and pushing a secondary cleaning solution into the sample cavity;

s108, introducing air from the sixth air channel (A6), and sequentially pushing the secondary cleaning solution in the sample cavity into the magnetic bead capture cavity (106) and the waste liquid cavity (105);

and S109, continuously introducing air from the sixth air channel (A6) to blow dry the magnetic beads in the magnetic bead capturing cavity (106).