WO2019006978A1 - 单通道化学发光微流控芯片及其检测方法 - Google Patents

单通道化学发光微流控芯片及其检测方法 Download PDF

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Publication number
WO2019006978A1
WO2019006978A1 PCT/CN2017/114692 CN2017114692W WO2019006978A1 WO 2019006978 A1 WO2019006978 A1 WO 2019006978A1 CN 2017114692 W CN2017114692 W CN 2017114692W WO 2019006978 A1 WO2019006978 A1 WO 2019006978A1
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Prior art keywords
liquid
whole blood
chip
chamber
quantitative
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PCT/CN2017/114692
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English (en)
French (fr)
Inventor
许行尚
陈⋅杰弗瑞
王龙
孙威严
万其露
杨申
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南京岚煜生物科技有限公司
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Application filed by 南京岚煜生物科技有限公司 filed Critical 南京岚煜生物科技有限公司
Priority to SG11201914032VA priority Critical patent/SG11201914032VA/en
Priority to EP17896325.2A priority patent/EP3457137B1/en
Priority to US16/079,440 priority patent/US10648975B2/en
Publication of WO2019006978A1 publication Critical patent/WO2019006978A1/zh

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    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558—Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G01N33/54386—Analytical elements
    • G01N33/54387—Immunochromatographic test strips
    • G01N33/54388—Immunochromatographic test strips based on lateral flow
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B—PERFORMING OPERATIONS; TRANSPORTING
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    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502715—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by interfacing components, e.g. fluidic, electrical, optical or mechanical interfaces
    • F—MECHANICAL ENGINEERING; LIGHTING; HEATING; WEAPONS; BLASTING
    • F16—ENGINEERING ELEMENTS AND UNITS; GENERAL MEASURES FOR PRODUCING AND MAINTAINING EFFECTIVE FUNCTIONING OF MACHINES OR INSTALLATIONS; THERMAL INSULATION IN GENERAL
    • F16K—VALVES; TAPS; COCKS; ACTUATING-FLOATS; DEVICES FOR VENTING OR AERATING
    • F16K99/00—Subject matter not provided for in other groups of this subclass
    • F16K99/0001—Microvalves
    • F16K99/0034—Operating means specially adapted for microvalves
    • F16K99/0042—Electric operating means therefor
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/52—Use of compounds or compositions for colorimetric, spectrophotometric or fluorometric investigation, e.g. use of reagent paper and including single- and multilayer analytical elements
    • G01N33/525—Multi-layer analytical elements
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54366—Apparatus specially adapted for solid-phase testing
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53—Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/543—Immunoassay; Biospecific binding assay; Materials therefor with an insoluble carrier for immobilising immunochemicals
    • G01N33/54393—Improving reaction conditions or stability, e.g. by coating or irradiation of surface, by reduction of non-specific binding, by promotion of specific binding
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2200/00—Solutions for specific problems relating to chemical or physical laboratory apparatus
    • B01L2200/14—Process control and prevention of errors
    • B01L2200/143—Quality control, feedback systems
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/046—Function or devices integrated in the closure
    • B01L2300/047—Additional chamber, reservoir
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/04—Closures and closing means
    • B01L2300/046—Function or devices integrated in the closure
    • B01L2300/048—Function or devices integrated in the closure enabling gas exchange, e.g. vents
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/0681—Filter
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/06—Auxiliary integrated devices, integrated components
    • B01L2300/069—Absorbents; Gels to retain a fluid
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0809—Geometry, shape and general structure rectangular shaped
    • B01L2300/0825—Test strips
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
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    • B01L2300/00—Additional constructional details
    • B01L2300/08—Geometry, shape and general structure
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0874—Three dimensional network
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
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    • B01L2300/08—Geometry, shape and general structure
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    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
    • B01L3/50—Containers for the purpose of retaining a material to be analysed, e.g. test tubes
    • B01L3/502—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures
    • B01L3/5027—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip
    • B01L3/502723—Containers for the purpose of retaining a material to be analysed, e.g. test tubes with fluid transport, e.g. in multi-compartment structures by integrated microfluidic structures, i.e. dimensions of channels and chambers are such that surface tension forces are important, e.g. lab-on-a-chip characterised by venting arrangements

Definitions

  • the invention relates to a single channel chemiluminescence microfluidic chip and a detection method thereof, and belongs to the technical field of immuno lateral chirp chromatography.
  • Immunological lateral tomography diagnostic techniques are suitable as a stable and practical technique for use in a variety of real-time tests (POCT) or in the field.
  • the coefficient of variation (CV) is large due to systemic reasons, and accurate quantification cannot be achieved.
  • the immunological diagnosis method based on microfluidic technology can effectively avoid the above problems.
  • Microfluidic is divided into passive and active. Passive microfluidics still require capillary forces to achieve lateral forward chromatography of the fluid. However, due to the different viscosity of different samples, especially whole blood samples, the liquid flow rate cannot be unified.
  • Active microfluidic can effectively avoid the above problems, can give forward thrust, make the liquid flow forward evenly, and avoid the difference of test values caused by different flow rates.
  • Active microfluidic power is driven by centrifugal force, electrowetting drive, pressure drive (electrolysis pump, compressed gas pump, chemical decomposition pump, direct differential pressure drive)
  • None of the patents 1), 2) and 3) relates to the monitoring of the exact position of the liquid as it flows through the chip.
  • the chips of the above three patents do not have fluid in the channel or cavity of the chip before the final test result. The filling behavior is monitored.
  • the labeled antibody and the coated antibody are placed in different cavities, which is disadvantageous to the simplification of the chip structure, and at the same time coated with the antibody and the labeled antibody. In the process of reaction, it can not be fully mixed, which affects the test efficiency.
  • the present invention is directed to the deficiencies of the prior art, and provides a single-channel chemiluminescence microfluidic chip, which subtly coats coated antibody and labeled antibody in the same reaction tank, simplifies the reagent operation process, and has physical physics between them.
  • the partition is effective to avoid non-specific binding.
  • by designing the special structure of the reaction tank when the fluid passes through the reaction tank, a liquid flow similar to "wave" can be generated, which promotes the mixed mixing of the coated antibody and the labeled antibody, thereby effectively improving the test efficiency and sensitivity.
  • a single-channel chemiluminescence microfluidic chip comprises a chip body, the chip body comprises an optical path scanning window, a whole blood filter cavity, a quantitative-reaction cavity and a waste liquid cavity, and a liquid outlet of the whole blood filter cavity passes through the microfluidic flow channel It is connected with the quantitative-reaction chamber and the waste liquid chamber in turn;
  • the optical path scanning window is set above the quantitative-reaction chamber, and the inlet port of the quantitative-reaction chamber can pass through the cleaning liquid delivery branch, the color-developing liquid delivery branch and the corresponding
  • the external liquid passage is connected, and the whole blood filter chamber can be connected with the external air passage;
  • the quantitative-reaction chamber is divided into a lower quantitative-reaction tank and a reaction tank capable of sealing the pool of the quantitative-reaction tank a cover plate;
  • the dosing-reaction cell is divided into three compartments by perpendicular to the flow of the fluid to the two partition plates a, wherein the labeled cells are placed in the middle compartment,
  • the liquid outlet of the whole blood filter chamber is connected to the liquid inlet of the quantitative-reaction chamber through the first backflow prevention device, and the liquid outlet of the cleaning liquid delivery branch and the color developing liquid are transported.
  • the liquid outlet of the branch is gathered, it is connected to the inlet of the quantitative-reaction tank through the second backflow prevention device;
  • the first backflow prevention device and the second backflow prevention device have the same structure, and both include an anti-reflux structure and anti-reflux
  • the structure is located above the microfluidic flow channel and has a backflow prevention channel;
  • the anti-reflux flow channel can raise the liquid level of the microfluidic flow channel at the installation position of the anti-reflux structure; when the microfluidic flow channel is connected to the external gas path Under the air pressure provided by the external air passage, the pressure of the liquid level of the microfluidic flow passage raised by the backflow prevention passage is overcome, and the fluid in the microfluidic flow passage on both sides of the backflow prevention device is caused to flow
  • the microfluidic flow path between the quantitative-reaction chamber and the waste liquid chamber A valve device is installed, the valve device includes a detecting mechanism for sensing whether a fluid flows through a valve device installation position; a valve device in a normally open state in the chip body, and a value fed back by the detecting mechanism indicates that the fluid flows through the valve device
  • the position is installed, it can be automatically closed and in the closed state; the valve device in the closed state can be opened regularly and in circulation; the valve device in the circulation state can be circulated in the valve device under the air flow provided by the external air passage.
  • the first absorbent paper is placed in the waste liquid chamber, and the waste liquid chamber has two connected waste liquid chamber bodies, respectively, a waste liquid chamber split a, and a waste liquid chamber split body. b, wherein: an inner air wall of the waste liquid chamber a and the first absorbent paper can form an air flow passage, and the waste liquid chamber split b has an air outlet.
  • the waste liquid chamber comprises a waste liquid pool, a middle cover plate capable of sealing the pool opening of the waste liquid pool, and an upper cover plate capable of covering the middle cover plate, wherein the waste liquid pool is separated from the waste liquid chamber
  • the corresponding wall of the body a is provided with a plurality of waste liquid guiding members, and a paper suction fixing column is installed at the bottom of the waste liquid pool close to the waste liquid guiding member; the waste liquid guiding member has a curved tooth shape
  • Each waste liquid guiding member is arranged in turn along the wall of the waste liquid pool according to its own arc shape, and the first absorbent paper is placed in the waste liquid pool through the paper suction fixing column, and the side of the first absorbent paper is The tip of the waste deflector touches.
  • the middle cover plate is provided with a middle layer through hole penetrating the pool opening of the waste liquid pool at a portion corresponding to the waste liquid chamber body a, and a pool mouth with the waste liquid pool is disposed at a position corresponding to the waste liquid chamber body b a continuous middle vent hole, and a water absorbing paper groove is disposed at a position of the middle vent hole, wherein the second absorbent paper is placed in the absorbing paper recess; the upper cover is provided at a position corresponding to the middle through hole
  • the vent hole sealed by the middle vent hole is capped, and an upper vent hole communicating with the middle vent hole is provided at a position corresponding to the middle vent hole.
  • the whole blood filter chamber has a sample portion connectable with an external gas, and the sample portion has a sample hole and a vent hole; the distance of the vent hole relative to the bottom of the whole blood filter chamber is larger than the sample hole The spacing from the bottom of the whole blood filtration chamber.
  • the injection portion has an annular groove and an annular flange, the outer side wall of the annular flange is an inner side wall of the annular groove, and the air source interface is disposed in the annular groove;
  • the vent hole includes the first ventilation a first venting hole is disposed at an end surface of the annular flange, the annular flange is engaged with the sampling hole through a converging transition surface, and the second venting hole is disposed adjacent to the inner wall of the annular flange to be in a convergence transition And the second venting hole is located adjacent to the position of the first venting hole.
  • the whole blood filter chamber comprises a whole blood filter sample tank and a top cover covering the whole blood filter sample pool, and the injection portion is disposed on the top cover;
  • the whole blood filter sample pool has a filter sample outlet Whole blood filter pool
  • the side wall extends in a tapered manner to the outlet of the filter sample, the side wall extending in a tapered manner is provided with a toothed arrangement with drainage;
  • the whole blood filter sample pool is covered with whole blood filter paper, and all The sides of the blood filter paper can be in contact with the inwardly convex tips of the teeth of the adjacent sidewalls of the whole blood filter sample cell.
  • Another technical object of the present invention is to provide a single-channel chemiluminescence microfluidic chip, comprising a chip body, the chip body having a three-piece structure, comprising an upper layer chip, a middle layer chip and a lower layer chip stacked in this order from top to bottom;
  • the upper layer chip, the middle layer chip and the lower layer chip are mutually connected by means of a positioning column and a positioning hole, and the layers are mutually connected;
  • the lower layer chip is provided with a whole blood filter tank, a quantitative-reaction tank and a waste liquid pool, and the liquid outlet of the whole blood filter tank is sequentially connected to the quantitative-reaction tank and the waste liquid pool through the microfluidic flow passage, and at the same time
  • the inlet port of the quantitative-reaction cell can be respectively connected to the corresponding external liquid path through the cleaning liquid delivery branch and the coloring liquid delivery branch, and the whole blood filtration pool can be connected with the external gas path;
  • the quantification-reaction cell is divided into three compartments by perpendicular to the flow of the fluid to the two partitions a arranged, wherein the labeled cells are placed in the compartment in the middle, and the compartments on both sides are placed. Coating the antibody; the reaction cell cover plate is divided into two cover plates by a vertical direction perpendicular to the flow direction of the partition plate b facing the quantitative-reaction cell, and each cover plate is filled in the split body Coated antibody;
  • the middle layer chip is provided with a sample injection site at a position corresponding to the whole blood filter pool, and a reaction cell cover plate capable of sealing the pool opening of the quantitative-reaction cell is disposed at a position corresponding to the quantitative-reaction cell, At the corresponding position of the liquid pool, a middle cover plate capable of sealing the pool opening of the waste liquid pool is provided, and a cleaning liquid interface is respectively disposed at a position corresponding to the liquid inlet of the cleaning liquid delivery branch and the liquid inlet of the color developing liquid delivery branch. , color developing fluid interface;
  • the upper layer chip is provided with an injection portion through hole at a position corresponding to the injection portion, an optical path scanning window is disposed at a position corresponding to the reaction cell cover plate, and an upper cover plate is disposed at a position corresponding to the middle cover plate.
  • a first backflow prevention device is installed on the microfluidic flow channel between the whole blood filter tank and the quantitative-reaction cell, and the microfluid between the cleaning liquid transport branch and the color developing liquid transport branch and the quantitative-reaction cell Installing a second backflow prevention device on the control flow channel;
  • the first backflow prevention device and the second backflow prevention device have the same structure, and each comprises an anti-reflux structure, the anti-reflux structure is located above the microfluidic flow channel, and has a backflow prevention channel; and the anti-reflux flow channel can lift the anti-reflux structure.
  • a valve device is mounted on the microfluidic flow path between the quantitative-reaction cell and the waste liquid pool, the valve device includes a detecting mechanism for sensing whether the fluid flows through the valve device installation position; and is normally open in the chip body
  • the valve device the value reported by the detecting mechanism indicates that the fluid can be automatically closed when the fluid flows through the valve device installation position, and is in a closed state; the valve device in the shut-off state can be opened periodically and in a flowing state; the valve device in the circulation state is in The fluid can be circulated in the valve device by the air flow provided by the external air circuit.
  • a further technical object of the present invention is to provide a method for detecting a single-channel chemiluminescence microfluidic chip, comprising the following steps:
  • Step 1 Add a whole blood sample to the sample well of the whole blood filter chamber;
  • Step 2 After the chip body is incubated for a period of time t1, the chip body is in contact with the external gas path, the external liquid path and the pressing mechanism, so that the cleaning liquid conveying branch of the chip body, the coloring liquid conveying branch and the corresponding external liquid The road is connected, and the whole blood filter chamber is connected with the external gas path; at the same time, in the valve device, the lower pressing head of the pressing mechanism is in contact with the inner sealing film of the conductive sealing ring, and the capacitance probe and the capacitive probe contact of the conductive sealing ring are touch;
  • Step 3 The external gas path is slowly inflated to the chip body, and the whole blood sample is filtered under the filtration of the whole blood filter paper to become plasma and enter the whole blood filter chamber;
  • Step 4 Under the continuous air pressure of the external airway, the plasma in the whole blood filtration chamber continues to flow along the microfluidic flow channel, sequentially flows through the first backflow prevention device, the quantitative-reaction chamber, and flows in.
  • the valve device at this time, once the plasma contacts the conductive sealing ring of the valve device, the capacitance probe senses the capacitance of the capacitance probe contact of the conductive sealing ring that contacts the contact, changes the valve device, and closes the microfluidic flow channel, At the same time, the external air circuit is closed and the pressurization is stopped;
  • Step 5 separating the main body of the chip from the external air path, the external liquid path and the pressing mechanism; and mixing the plasma in the quantitative-reaction chamber with the coated antibody and labeled antibody embedded therein for 3-10 minutes for performing an immune reaction ;
  • Step 6 contacting the chip body with the external gas path, the external liquid path and the pressing mechanism, so that the cleaning liquid conveying branch of the chip body and the coloring liquid conveying branch are connected with the corresponding external liquid path, and the whole blood filtering chamber is It is connected with the external gas path; at the same time, in the valve device, the lower pressing head of the pressing mechanism and the inner sealing of the conductive sealing ring are closed When the membrane touches, the capacitance probe touches the capacitance probe contact of the conductive sealing ring;
  • Step 7 Start the external gas path, and the air pressure provided by the external gas path will push the plasma in the chip body to flow along the microfluidic flow channel until the sample hole and the plasma in the microfluidic flow channel are blown. At this time, the capacitance probe The sensed capacitance value returns to the initial value, and the external air path is closed;
  • Step 8 Starting the cleaning liquid device in the external liquid path, causing the cleaning liquid to sequentially flow through the second anti-backflow device and the quantitative-reaction chamber installed on the cleaning liquid delivery branch, and then arrive at the valve device; once the cleaning liquid contacts the valve device
  • the conductive sealing ring causes the capacitance probe to sense the capacitance of the capacitive probe contact of the conductive sealing ring that contacts it, touches the valve device, closes the microfluidic flow path, and at the same time, closes the external pneumatic circuit, stops the pressing ;
  • Step IX the cleaning solution is mixed in the chip body, especially the quantitative-reaction chamber for 1-3 minutes for cleaning;
  • Step 10 Start an external gas path, provide air pressure to push the cleaning liquid to move forward, and blow dry the liquid in the chip body. At this time, the capacitance value sensed by the capacitance probe returns to an initial value, and the external gas path is closed;
  • Step XI repeat step 8 to step 10 3-8 times;
  • Step 12 Start the color developing liquid device in the external liquid path, and prompt the color developing liquid to sequentially flow through the anti-backflow device and the quantitative-reaction chamber installed on the coloring liquid conveying branch, and then reach the valve device; the coloring liquid once contacted
  • the conductive sealing ring of the valve device causes the capacitance probe to sense the capacitance of the capacitive probe contact of the conductive sealing ring that is in contact with it, activates the valve device, closes the microfluidic flow channel, and at the same time, closes the external pneumatic circuit, Stop pressing;
  • Step 13 The color developing solution is mixed in the quantitative-reaction chamber for 3-8 minutes to carry out a color reaction;
  • Step 14 Separating the chip body from the external air path, the external liquid path, and the pressing mechanism to complete the detection.
  • the present invention has the following advantages over the prior art:
  • the present invention provides a specific structure of a quantitative-reaction chamber in which a coated antibody and a labeled antibody can be simultaneously placed, and at the same time, there is a physical partition between the coated antibody and the labeled antibody (separator a , the separator b, the labeled antibody placed in the reaction cell cover plate and the labeled antibody placed in the quantitative-reaction cell have a gap), thereby simplifying the reagent operation procedure and effectively avoiding the coating between the antibody and the labeled antibody.
  • Non-specific binding, in addition, the dosing-reaction chamber of the present invention, through the partition plate a and the baffle b provided, can make the flow flowing through the quantification-reaction chamber wavy, and the coating can be well coated.
  • the antibody and labeled antibody are mixed to effectively improve the test efficiency and sensitivity.
  • the front end of the inlet port of the quantitative-reaction chamber of the present invention is equipped with an anti-backflow device (first backflow prevention device and a second anti-backflow device), so that one device can operate a plurality of microfluidic chips to function as a multi-pass
  • an anti-backflow device first backflow prevention device and a second anti-backflow device
  • the rear end of the quantitative-reaction chamber of the present invention is equipped with a valve device, and the valve device and the device are closely combined, and the valve is closed at any time through the sealing ring with the sealing membrane and the pressing device on the device, and Can be reused.
  • the valve device does not require high production processes.
  • the timing of the flow of liquid in the flow channel to the sealing ring can be detected in time, and the sealing ring can be accurately closed, which can eliminate the air bubbles in front of the liquid and can also throttle the liquid, and functions as a semi-permeable membrane (breathable liquid-impermeable) device. Save the semi-permeable membrane device function module.
  • the whole blood filter chamber of the present invention is provided with a sample hole and a vent hole at the injection site to avoid the problem of blockage of the whole blood sample caused by the active microfluidic control; the vent hole is set to a height of the same
  • the second venting hole, the first venting hole, the spacing of the second venting hole relative to the bottom of the whole blood filtering chamber is larger than the spacing of the sampling hole relative to the bottom of the whole blood filter chamber, in particular, the first venting hole is set at a higher height than the second venting hole Hole, in the process of loading, even if the sample hole and the second vent hole are blocked, the whole blood filter sample chamber can be connected to the gas source through the first vent hole, and the active microfluidic gas path is circulated;
  • the present invention extends the side wall of the whole blood filter sample cell to the outlet position of the filter sample in a tapered manner, and can function as a drainage, and the side wall extending in a tapered manner has a tooth shape with drainage effect.
  • the setting can effectively avoid the problem that the gap (the gap between the side wall of the whole blood filter sample and the side of the whole blood filter paper) due to production error cannot completely filter the whole blood sample.
  • the invention provides a gas passage in the waste liquid chamber, and discharges the gas through the gas outlet port of the waste liquid chamber, thereby effectively ensuring that the liquid flow can be well absorbed by the absorbent paper to prevent backflow, splashing or liquid leakage of the waste liquid.
  • FIG. 1 is a schematic exploded view showing the single-channel chemiluminescence microfluidic chip of the present invention
  • FIG. 2 is a schematic structural view of a lower layer chip according to the present invention.
  • Figure 3 is a schematic enlarged view of the waste liquid pool portion (without absorbent paper) of Figure 2;
  • Figure 2 Figure 3: 3-1, whole blood filter pool; 3-1-1, sample flow guide; 3-2, quantitative-reaction tank; 3-2-1, partition a; 3-3, Waste liquid pool; 3-3-1, waste liquid guiding piece; 3-3-2, absorbent paper positioning column; 3-4, cleaning liquid conveying branch; 3-5, coloring liquid conveying branch; 6, the lower layer chip positioning port; 3-7, chip fixing column; 6, valve device; 9-a, first reflow device; 9-b, second reflow device;
  • Figure 4 is a schematic enlarged plan view of the waste liquid pool portion (with absorbent paper) of Figure 2;
  • Figure 5 is a schematic structural view of one direction (front side) of the middle chip
  • Figure 6 is a schematic structural view of the middle layer chip in the other direction (back side);
  • 2-1 sample loading tank
  • 2-1-1 venting hole
  • 2-1-2 sample loading hole
  • 2-2 chip positioning hole
  • 2-3 return hole of the first reflow device
  • -4 the return hole of the second reflow device
  • 2-5 the external interface of the cleaning liquid branch
  • 2-6 the external interface of the color developing liquid branch
  • 2-7 the mounting groove of the conductive sealing ring
  • 2-8 the middle layer through hole of the waste liquid pool
  • 2-9 the air outlet
  • 2-10 the reaction pool cover
  • Figure 7 is a schematic structural view of the front side of the upper chip
  • Figure 8 is a schematic structural view of the back side of the upper chip
  • Figure 7 and Figure 8 1-1, injection part through hole; 1-2, optical path scanning window; 1-3, effluent hole of waste liquid pool; 1-4, through hole of external interface of cleaning liquid; 1-5 , through hole of external connection port of color developing liquid; 1-6, capacitance probe through hole; 1-7, lower pressure head through hole;
  • Figure 9 is a schematic structural view of a three-piece backflow prevention structure
  • Figure 10 is a cross-sectional view of Figure 9;
  • Figure 11 is a schematic structural view of a second structure anti-reflux structure
  • Figure 12 is a cross-sectional view of Figure 11;
  • FIG. 13 is a schematic structural view of a lower layer chip in a backflow prevention structure of a third structural form
  • FIG. 14 is a schematic structural view of an upper layer chip in a backflow prevention structure of a third structural form
  • upper chip 1 middle chip 2; anti-backflow liquid outlet 2-a; anti-backflow communication channel 2-b; anti-reflux bump 2-c; anti-return flow channel 2-d; lower chip 3; microfluidic flow channel 3-a; microfluidic flow channel spacer 3-b; connecting column 3-c;
  • Figure 15 is a schematic exploded view of the valve device of the present invention.
  • Figure 16 is a schematic structural view of a middle chip at a corresponding position of the valve device
  • Figure 17 is a perspective view showing the structure of the conductive sealing ring
  • Figure 18 is a cross-sectional view of one direction of Figure 16;
  • Figure 19 is a cross-sectional view of the other direction of Figure 16;
  • Figure 15 - Figure 19 1, the upper chip; 2, the middle chip; 2-e, the middle chip flow channel; 2-f, the seal ring groove; 3, the lower chip; 6-1, the pressing mechanism; 6- 2, conductive sealing ring; 6-2-1, outer sealing ring; 6-2-2, internal sealing film; 6-2-3, capacitance probe contact; 6-3, detection probe.
  • spatially relative terms such as “above”, “above”, “on top”, “above”, etc., may be used herein to describe as in the drawings.
  • the exemplary term “above” can include both “over” and "under”.
  • the device can also be positioned in other different ways (rotated 90 degrees or in other orientations).
  • the invention discloses a single-channel chemiluminescence microfluidic chip, comprising a chip body, wherein the chip body comprises an optical path scanning window, a whole blood filter cavity, a quantitative-reaction cavity and a waste liquid cavity, and a liquid outlet of the whole blood filter cavity
  • the microfluidic flow channel is sequentially connected with the quantitative-reaction chamber and the waste liquid chamber;
  • the optical path scanning window is disposed above the quantitative-reaction chamber, and the inlet port of the quantitative-reaction chamber can respectively pass through the cleaning liquid to transport the branch and develop color
  • the liquid delivery branch is in communication with the corresponding external liquid path, and the whole blood filtration chamber is connectable with the external gas path;
  • the quantitative-reaction chamber is divided into a lower quantitative-reaction cell and a quantification-reaction cell a cell cover sealed by the cell;
  • the dosing-reaction cell is divided into three compartments by perpendicular to the flow of the two partitions a, wherein the labeled cells are placed in
  • reaction cell cover plate is divided into two cover plates by a vertical direction perpendicular to the flow direction of the partition plate b facing the quantitative-reaction cell, and each is divided into two cover plates, each The coated cover is filled with coated antibodies.
  • the liquid outlet of the whole blood filter chamber is connected to the liquid inlet of the quantitative-reaction chamber through the first backflow prevention device, and the liquid outlet of the cleaning liquid delivery branch and the liquid outlet of the color developing liquid delivery branch are gathered Thereafter, the second backflow prevention device is connected to the liquid inlet of the dosing-reaction cell; the first backflow prevention device and the second backflow prevention device have the same structure, both of which include an anti-reflux structure, and the anti-reflux structure is located in the microfluidic flow channel Above, there is a backflow prevention channel; the backflow prevention channel can raise the liquid level of the microfluidic flow channel at the installation position of the anti-reflux structure; when the microfluidic flow channel is connected to the external gas path, the external gas path is provided Under the pressure driving, the pressure of the liquid level of the microfluidic flow channel raised by the backflow prevention passage is overcome, and the fluid in the microfluidic flow passage on both sides of the backflow prevention device is caused to flow.
  • a valve device is mounted on the microfluidic flow passage between the quantitative-reaction chamber and the waste liquid chamber, and the valve device includes a detecting mechanism for sensing whether the fluid flows through the installation position of the valve device; and is normally open in the chip body
  • the value reported by the detecting mechanism indicates that the fluid can be automatically closed when the fluid flows through the valve device installation position, and is in a closed state; the valve device in the closed state can be opened periodically and in a flowing state; the valve device in the circulating state
  • the fluid can be circulated in the valve device by the air flow provided by the external air passage.
  • the first absorbent paper is disposed in the waste liquid chamber, and the waste liquid chamber has two connected waste liquid chamber splits, respectively, a waste liquid chamber split a, a waste liquid chamber split b, wherein: the waste liquid chamber An air flow passage can be formed between the inner wall of the split body a and the first absorbent paper, and the waste liquid chamber split b has an air outlet.
  • the waste liquid chamber includes a waste liquid pool, a middle cover plate capable of sealing the pool opening of the waste liquid pool, and an upper cover plate capable of covering the middle cover plate, and the waste liquid pool is disposed at a wall corresponding to the waste liquid cavity a
  • a paper suction fixing column is installed at the bottom of the waste liquid pool close to the waste liquid guiding member;
  • the waste liquid guiding member has a curved tooth shape, and each waste liquid guiding member According to its own arc shape, it is arranged in turn along the wall of the waste liquid pool, first The absorbent paper is placed in the waste liquid pool through the paper suction fixing column, and the side of the first absorbent paper is in contact with the tip end of the waste liquid guiding member.
  • the middle cover plate is provided with a middle layer through hole penetrating the pool opening of the waste liquid pool at a portion corresponding to the waste liquid chamber body a, and a pool mouth with the waste liquid pool is disposed at a position corresponding to the waste liquid chamber body b a communicating middle vent hole, and a water absorbing paper recessed groove is disposed at a position of the middle vent hole, wherein the first absorbent paper is disposed in the absorbing paper recess; the upper cover is disposed at a position corresponding to the middle through hole
  • the vent hole sealed by the middle vent hole is capped, and an upper vent hole communicating with the middle vent hole is provided at a position corresponding to the middle vent hole.
  • the whole blood filter chamber has an injection portion connectable with an external gas, and the injection portion has a sample hole and a vent hole; the distance of the vent hole relative to the bottom of the whole blood filter chamber is larger than the bottom of the whole blood filter chamber Pitch.
  • the injection portion has an annular groove and an annular flange, the outer side wall of the annular flange is an inner side wall of the annular groove, and the air source interface is disposed in the annular groove;
  • the venting hole comprises a first venting hole and a second venting hole;
  • the first venting hole is disposed at an end surface of the annular flange, the annular flange is engaged with the sampling hole through a converging transition surface, the second venting hole is disposed on the converging transition surface of the annular plenum against the inner wall of the annular flange, and the second venting hole The position is adjacent to the position of the first venting hole.
  • the whole blood filter chamber includes a whole blood filter sample pool and a top cover covering the whole blood filter sample pool, and the injection portion is disposed on the top cover;
  • the whole blood filter sample pool has a filter sample outlet, and the whole blood filter sample pool
  • the side wall extends in a tapered manner to the outlet of the filter sample, the side wall extending in a tapered manner is provided with a toothed arrangement with drainage;
  • the whole blood filter sample pool is provided with whole blood filter paper, and whole blood
  • the sides of the filter paper can be in contact with the indented tips of the teeth of the toothed sidewalls of the adjacent whole blood filter cell.
  • the single-channel chemiluminescence microfluidic chip of the present invention comprises a chip body, and the chip body has a three-piece structure, including an upper layer chip, a middle layer chip and a lower layer which are sequentially stacked from top to bottom.
  • the chip; the upper layer chip, the middle layer chip and the lower layer chip are respectively connected by a positioning column and a positioning hole, and the layers are mutually connected;
  • the lower layer chip as shown in FIG. 2 and FIG. 3, is provided with a whole blood filter tank, a quantitative-reaction tank and a waste liquid pool, and the liquid outlet of the whole blood filter tank is sequentially passed through the microfluidic flow channel and the quantitative-reaction tank
  • the waste liquid pool is connected, and the liquid inlet of the quantitative-reaction tank can be connected to the corresponding external liquid path through the cleaning liquid delivery branch and the color development liquid delivery branch, respectively, and the whole blood filtration pool can be connected with the external gas.
  • the road is connected.
  • the whole blood filter sample cell has a filter sample outlet, and a side wall of the whole blood filter sample cell is extended in a tapered manner to a filter sample outlet position, and the side wall extending in a tapered manner has a toothed arrangement with drainage effect
  • the whole blood filter sample pool is provided with whole blood filter paper, and the sides of the whole blood filter paper can be in contact with the convex tips of the teeth of the toothed side walls of the adjacent whole blood filter sample pool.
  • the whole blood filter paper is pressed at the bottom of the whole blood filter sample pool through the filter paper pressure point set at the lower end of the injection portion.
  • the quantification-reaction cell is divided into three compartments by perpendicular to the flow of the fluid to the two partitions a arranged, wherein the labeled cells are placed in the compartment in the middle, and the compartments on both sides are placed
  • the coated cell cover plate is divided into two cover plates separated by a vertical direction perpendicular to the fluid flow direction on the plate surface facing the quantitative-reaction cell, and each cover plate is buried in the split body. Filled with coated antibodies.
  • the waste liquid pool as shown in FIG. 4, has a connected waste liquid pool split a, a waste liquid pool split b, and a waste liquid guide body is provided on the wall of the waste liquid pool split a, and is tight
  • a paper suction fixing column is installed at the bottom of the waste liquid guiding member; the waste liquid guiding member has a curved tooth shape, and each waste liquid guiding member rotates along the arc according to its own arc shape along the waste liquid pool.
  • the pool walls of a are arranged in sequence, and the first absorbent paper is placed in the waste liquid pool through the paper suction fixing column, and the side of the first absorbent paper is in contact with the tip end of the waste liquid guiding member.
  • the middle layer chip as shown in FIG. 5 and FIG. 6, is provided with a sample injection site at a position corresponding to the whole blood filter pool, and is provided at a position corresponding to the quantitative-reaction cell to seal the pool port of the quantitative-reaction cell
  • the reaction cell cover plate is provided with a middle cover plate capable of sealing the pool port of the waste liquid pool at a position corresponding to the waste liquid pool, and the liquid inlet port of the cleaning liquid delivery branch and the liquid supply port of the color development liquid delivery branch Corresponding positions are respectively provided with a cleaning liquid interface and a color developing liquid interface;
  • the middle layer cover is provided with a through-medium through hole at a position corresponding to the waste liquid pool body a, at a position corresponding to the waste liquid pool split b
  • a middle vent hole is disposed at the middle, and a water absorbing paper groove is disposed at a position of the middle vent hole, and a second absorbent paper is disposed in the absorbing paper recess.
  • the injection portion has an annular groove and an annular flange, the outer side wall of the annular flange is an inner groove wall of the annular groove, the annular flange is disposed on the upper surface of the middle chip, and the outer groove wall of the annular groove is disposed on the upper chip
  • the middle chip is provided with an annular groove groove bottom between the outer groove wall of the annular groove and the outer side wall of the annular flange, and the air source interface is disposed in the annular groove;
  • the vent hole includes the first vent hole and the second ventilation a first venting hole is disposed at an end surface of the annular flange, the annular flange is engaged with the sampling hole through a tapered transition surface, and the second venting hole is disposed on the tapered transition surface against the inner wall of the annular flange.
  • the upper layer chip as shown in FIG. 7 and FIG. 8, is provided with a through-hole of the injection portion at a position corresponding to the injection portion, and an optical path scanning window is disposed at a position corresponding to the reaction cell cover, and the middle cover is disposed at the position corresponding to the reaction cell cover.
  • the upper cover is disposed at the position, and the cleaning liquid interface through hole and the color developing liquid interface through hole are respectively disposed at positions corresponding to the cleaning liquid interface and the color developing liquid interface; the upper cover plate is located at a position corresponding to the middle through hole
  • a vent cover capable of sealing the middle vent hole is provided, and an upper vent hole communicating with the middle vent hole is provided at a position corresponding to the middle vent hole.
  • a first anti-reflux device is installed on the microfluidic flow channel between the whole blood filter tank and the quantitative-reaction cell, and the cleaning liquid transport branch and the color developing liquid transport branch are concentrated, and quantitatively- A second backflow prevention device is installed on the microfluidic flow path between the reaction cells.
  • the first backflow prevention device and the second backflow prevention device have the same structure, as shown in FIG. 9-14, each includes an anti-reflux structure, and the anti-reflux structure is located above the microfluidic flow channel, and has a backflow prevention channel;
  • the flow channel can raise the liquid level of the microfluidic flow channel at the installation position of the anti-reflux structure; when the microfluidic flow channel is connected to the external gas path, the anti-return flow path is lifted under the air pressure provided by the external gas path
  • the pressure of the liquid level of the microfluidic flow channel causes the fluid in the microfluidic flow channel on both sides of the backflow prevention device to be in a flowing state.
  • the extending direction of the anti-reflux flow passage is perpendicular to the extending direction of the microfluidic flow passage at the installation position of the anti-reflux structure; and the microfluidic flow passage at the installation position of the anti-reflux structure is divided into two by the microfluidic flow passage spacer
  • the segments are respectively a microfluidic inlet flow channel and a microfluidic fluid outlet flow channel; the upper end of the anti-reflux flow channel is closed, and the lower end is bridged over the microfluidic flow channel spacer, and respectively is connected with the microfluidic flow inlet The flow channel and the microfluidic fluid outlet are connected.
  • the anti-reflux flow channel has anti-reflux bumps, and two anti-reflux communication channels are formed between the anti-reflux bump and the anti-return chute; the two anti-reflux connecting flow channels are respectively microfluidic The inlet flow passage and the microfluidic liquid outlet flow passage are connected; the lower end of the anti-reflux projection is flush with the lower end of the anti-return flow passage, and the upper end of the anti-reflux projection is disposed lower than the upper end of the anti-return flow passage.
  • the anti-reflux bump is disposed in a trapezoidal shape in a cross section between the two backflow prevention communication passages.
  • the microfluidic channel spacer is a wedge block
  • the backflow prevention channel is a wedge groove having a shape similar to that of the microfluidic channel spacer.
  • the microfluidic flow channel at the installation position of the anti-reflux structure is symmetrically arranged on both sides of the microfluidic flow channel spacer with a wedge-shaped connecting column, and the anti-return flow channel has a wedge-shaped connecting groove that is coupled with the wedge-shaped connecting column.
  • a valve device is mounted on the microfluidic flow path between the quantitative-reaction cell and the waste liquid pool, and the valve device, as shown in FIGS. 15-19, includes a sensor device for sensing whether the fluid flows through the valve device.
  • the detecting mechanism of the mounting position; the valve device in the normally open state in the chip body, the value fed back by the detecting mechanism indicates that the fluid can automatically close when the fluid flows through the valve device mounting position, and is in a closed state;
  • the valve device can be opened at a time and in a circulating state; the valve device in a circulating state can be circulated in the valve device by the air flow provided by the external air passage.
  • the valve device is a backflow prevention microvalve with anti-backflow function; and comprises a pressing mechanism, a detecting mechanism, a sealing member and a microvalve anti-reflux structure; wherein:
  • the pressing mechanism includes a lower pressing head located above the inner sealing film.
  • the detecting mechanism comprises a detecting probe connected in linkage with the lower pressing head and located above the probe contact, and the spacing between the detecting probe and the probe contact is smaller than the spacing between the lower pressing head and the inner sealing film .
  • the sealing member is a conductive sealing ring, including an outer sealing ring, an inner sealing film and a capacitance probe contact; the inner sealing film is in the inner ring of the outer sealing ring, and the capacitance probe contact is connected to the outer sealing ring.
  • the microvalve anti-reflux structure is arranged separately from the microfluidic flow channel at the installation position of the anti-backflow microvalve, and includes a through micro-valve anti-reflux flow passage;
  • the microfluidic flow channel at the installation position of the anti-backflow microvalve is divided into a micro valve inlet section and a microvalve outlet section by a microfluidic flow channel;
  • the microvalve inlet section has a liquid outlet, and the microvalve outlet The section has a liquid inlet;
  • microvalve anti-reflux flow channel is placed above the microfluidic flow channel traverse, and the microvalve anti-reflux flow channel can respectively correspond to the liquid inlet of the micro valve inlet section and the liquid inlet of the microvalve outlet section.
  • the inner cavity of the microvalve anti-reflux flow passage has a microvalve anti-reflux projection; the outer ring of the microvalve anti-reflux projection and the inner cavity of the microvalve anti-return flow passage are formed for mounting the outer sealing ring.
  • the sealing ring of the micro-valve has two through-flow passages, which are respectively a micro-valve anti-reflux inlet flow passage and a micro-valve anti-reflux outlet flow passage;
  • the micro-valve anti-reflux inlet flow passage is at the lower end and The liquid outlet of the micro valve inlet section is connected, and the lower end of the microvalve anti-reflux outlet passage is connected with the liquid inlet of the microvalve outlet section;
  • the outer sealing ring is embedded in the sealing groove of the sealing ring, and the inside is closed
  • the membrane can cover the upper end of the micro valve anti-reflux inlet flow channel and the upper end of the microvalve anti-reflux liquid flow channel;
  • the inner sealing membrane can simultaneously cover the liquid outlet of the micro valve inlet section and the liquid inlet of the microvalve outlet section, and is sealed by the outer sealing ring.
  • the detecting probe of the detecting mechanism touches the contact of the capacitor probe with the pressing action of the pressing mechanism; and detects whether the liquid flows into the valve at any time. At this time, the backflow prevention microvalve is open. In fact, in the case where there is a gap between the lower pressing head of the pressing mechanism and the inner closing film, the fluid can be driven by the air flow due to the deformation function of the inner sealing film itself and the deformation space at the mounting position. Microvalve In the liquid inlet section, after the deformation of the inner sealing membrane is overcome, the micro valve outlet section is entered.
  • the liquid When the gas path is turned on, the liquid is moved forward by the air pressure provided by the gas path, and the fluid in the liquid inlet section of the micro valve flows through the microvalve anti-reflux flow path to the micro valve outlet section, and the detection mechanism
  • the capacitance probe detects that the liquid flows over, and triggers the pressing mechanism 1 again.
  • the inner sealing membrane Under the driving of the lower pressure driving, the inner sealing membrane seals the upper end of the anti-reflux flow passage of the micro valve, and at this time, the anti-reflux micro valve is in a closed state;
  • the detecting mechanism comprises a capacitance probe, and the capacitive probe can be electrically connected to the capacitive probe contact under the driving of the pressing mechanism of the pressing mechanism.
  • the pressing mechanism stops downward, and at this time, between the lower pressing head and the inner sealing film There is a gap; when there is a change in the data sensed by the detecting probe, it indicates that there is a microfluidic flow path at the installation position of the valve device, triggering the action of the pressing mechanism, pushing the lower pressing head to continue downward until the lower pressing head is pressed
  • the inner sealing membrane is used to simultaneously seal the liquid outlet of the micro valve inlet section and the liquid inlet of the micro valve outlet section, and the valve device is in a closed state; when the lower pressing head of the pressing mechanism and the inner sealing membrane exist During the gap, the valve device is in an open state, and the internal sealing membrane has a deformation characteristic to ensure that fluid can flow out from the liquid outlet of the micro valve inlet section and from the inlet port of the microvalve outlet section.
  • the valve device has a three-piece structure, including an upper layer chip, a middle layer chip and a next chip which are sequentially distributed from top to bottom; the upper layer chip, the middle layer chip and the next chip are mutually coupled to each other; the micro valve The anti-reflux structure is disposed on the middle layer chip, and the microfluidic flow channel is disposed on the lower layer chip.
  • the recessed groove is disposed on the upper surface of the middle layer chip corresponding to the micro valve anti-reflux structure, and the sealing member is pressed in the embedded groove through the upper layer chip, and the upper layer chip Corresponding to the position of the inner sealing ring and the detecting contact respectively, the pressing head through hole and the probe passing hole are respectively opened; and the pressing mechanism is located above the upper layer chip, and the pressing mechanism is pressed down, and the lower pressing head is driven through the pressing head through the hole. Pressing the inner sealing membrane to seal the upper end of the microvalve anti-reflux flow passage.
  • the anti-return microvalve is closed; the capacitive probe moves toward the upper chip as the pressing mechanism 1 is pressed down until passing through the upper layer After the probe on the chip passes through the hole, it is in contact with the probe contact of the conductive sealing ring placed in the middle chip slot to realize electrical connection.
  • the invention creatively designs the valve device to fuse with the microfluidic liquid control backflow prevention device, saving space for the microfluidic chip.
  • the position space of the microfluidic chip is very limited, and the invention can effectively save the microfluidic flow channel space, and can make the flow path more effective. Avoid the problem that the liquid flow path is not tightly closed due to the short distance between the respective flow paths, and the flow path is penetrated.
  • the invention utilizes the conductivity of the conductive rubber material, combines the capacitance detecting probe of the device, observes the accurate timing of the liquid flowing into the valve in the flow channel, and closes the valve in time. Save semi-permeable membrane device.
  • the present invention provides the following detection method, which specifically includes the following steps:
  • Step 1 Insert the reagent chip into the instrument and read the above information
  • Step 2 pipette loading, 100 ul of whole blood is added to the sample well;
  • Step 3 the chip enters the inside of the instrument and is incubated for 2 minutes;
  • Step 4 The instrument is combined with the contact device of the chip, and includes a pneumatic device, two liquid path devices, and a valve pressure point with a conductive rubber probe;
  • Step 5 the pneumatic device is slowly inflated, and the whole blood sample is moved forward. During this process, the red blood cells are trapped by the blood filter paper, but due to the gas pressure, the red blood cells are clogged, and the sample hole may be poorly ventilated. Pushing the sample forward by the vent opening;
  • Step 6 After the sample passes through the blood filter paper, it becomes plasma, enters the flow channel, and is pushed by the pressure to pass through the backflow prevention device, and then the sample passes through the liquid flow channel, but since the liquid flow path terminal has the liquid path device pressing the chip, the liquid It is not possible to enter this channel, and the liquid flow path and sample flow direction are also different;
  • Step 7 The sample continues to move forward into the reaction quantification hole
  • Step 8 The sample continues to move forward and enters the valve device. Once the sample contacts the conductive rubber in the valve, the capacitance change will touch the valve switch and close the flow channel valve. At this time, there is a backflow prevention device behind the valve to prevent the valve from closing. The problem of liquids blowing away quickly. At the same time, the air circuit switch is closed and the pressure is stopped;
  • Step 9 The instrument and the chip contact device are separated, mixed for 3-10 minutes, and an immune reaction is performed;
  • Step 10 the instrument and the chip contact device are combined, the gas pushes the sample to move forward, blows the sample hole and the flow channel sample, and at this time, the conductive rubber probe data returns to the original value, and the gas path device is closed;
  • Step 11 the cleaning liquid path device is started, enters the flow channel, and then pushes through the anti-backflow device by pressure, and then the cleaning liquid passes through the sample flow channel, but the liquid cannot enter the channel because the sample flow channel terminal has the pneumatic device pressing the chip. And the flow direction of the liquid flow path and the sample are also different.
  • the other liquid path channel has a liquid path device pressing the chip at the end of the liquid flow path, and the liquid cannot enter the channel, and the liquid flow path and the sample flow direction are also different;
  • Step 12 the cleaning liquid continues to move forward into the reaction quantification hole
  • Step 13 The cleaning liquid continues to move forward and enters the valve device. Once the cleaning fluid contacts the conductive inside the valve Rubber, the change of capacitance will touch the valve switch and close the flow channel valve. At this time, there is a backflow prevention device behind the valve to avoid the problem that the valve is not closed in time and the liquid is quickly blown away. At the same time, the air circuit switch is closed and the pressure is stopped;
  • Step 14 mix for 1-3 minutes, and wash
  • Step 15 The gas pushes the cleaning liquid to move forward, and blows the sample hole and the flow channel liquid. At this time, the conductive rubber probe data returns to the original value, and the gas path device is closed;
  • Step 16 repeat steps 11 to 15 steps 3-8 times;
  • Step 17 the color liquid liquid circuit device is started, enters the flow channel, and then pushes through the anti-backflow device by pressure, and then the cleaning liquid passes through the sample flow channel, but the liquid cannot enter because the sample flow channel terminal has the pneumatic device pressing the chip.
  • This channel also has different flow directions for the liquid flow path and sample.
  • the other liquid path channel has a liquid path device pressing the chip at the end of the liquid flow path, and the liquid cannot enter the channel, and the liquid flow path and the sample flow direction are also different;
  • Step 18 the color developing solution continues to enter the reaction quantification hole
  • Step 19 The cleaning liquid continues to move forward and enters the valve device. Once the cleaning fluid contacts the conductive rubber in the valve, the capacitance change will touch the valve switch and close the flow channel valve. At this time, there is a backflow prevention device behind the valve to prevent the valve from closing. Not timely, the liquid quickly blows away the problem. At the same time, the air circuit switch is closed and the pressure is stopped;
  • Step 20 mixing for 3-8 minutes, performing a color reaction
  • Step 21 separating the instrument and the chip contact device
  • Step 22 the instrument reads data
  • Step 23 The instrument exits the chip and completes the test.

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Abstract

一种单通道化学发光微流控芯片及其检测方法，包括芯片本体，芯片本体包括光路扫描窗口（1-2）、全血过滤腔、定量-反应腔以及废液腔；定量-反应腔分体设置成处于下方的定量-反应池（3-2）以及能够将定量-反应池（3-2）的池口封接的反应池盖板（2-10）；定量-反应池（3-2）通过垂直于流体流向所布置的两块隔板a （3-2-1）等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；反应池盖板（2-10）在面向定量-反应池（3-2）的板面通过垂直于流体流向所布置的隔板b等分成两块盖板分体，每块盖板分体中均埋填有包被抗体。该芯片能够简化试剂操作流程，有效避免非特异性结合。此外，定量-反应腔的结构使得流体经过定量-反应腔时，能够产生类似于"波浪"的液流，促使包被抗体、标记抗体充分混合，有效地提高测试效率和灵敏度。

Description

单通道化学发光微流控芯片及其检测方法 技术领域
本发明涉及一种单通道化学发光微流控芯片及其检测方法，属于免疫侧向层析诊断技术领域。
背景技术
免疫侧向层析诊断技术作为一种稳定和实用的技术适合在多样的即时检验(POCT)或者现场使用。
在免疫层系反应系统中，由于系统原因导致变异系数CV(Coefficient of Variation)大，无法达到精确定量。而基于微流控技术的免疫诊断方法，可以有效的避免上述问题。
微流控又分被动式和主动式两种。被动式微流控还是需要毛细血管力来达到液体向前的侧向层析。但是由于不同样本特别是全血样本的粘稠度不同，导致液体流速无法统一。
主动式微流控可以有效避免上述问题，可以给向前的推力，使液体均匀的向前流动，避免因为不同流速导致的测试值差异。
主动式微流控的动力有离心力驱动、电润湿驱动、压力驱动(电解泵、压缩气体泵、化学分解泵、直接气压差驱动)
但是如果要达到随意控制液体速度的目的，不但要有推动力，还要有阀门控制，还要有防回流免得液体因为压力去除，回流回去。
现有相关技术，芯片的制作以及芯片的应用，请参考以下专利：
1)CN203899622U一种微流控芯片
2)CN106353491A微流控床旁边快速诊断试剂盒
3)CN205941345U用于生物检测的微流控芯片
其中，专利2)和3)中，未对加入到芯片中的样本液体进行定量，而要实现定量检测，必须实现对加入的样本的定量和预先放置于芯片内的试剂的定量。
专利1)，2)和3)中都未涉及液体在芯片中流动时准确位置的监测，换句话说，上述3项专利的芯片在最后的检测结果之前没有对流体在芯片内通道或腔体的填充行为做监测。
另外，现有技术中，为防止标记抗体、包被抗体的非特异性结合，标记抗体和包被抗体是置于不同的腔体中，不利于芯片结构的简化，同时包被抗体与标记抗体的在反应过程中也不能得到充分混合，影响测试效率。
发明内容
本发明针对现有技术的不足，提供一种单通道化学发光微流控芯片，其将包被抗体和标记抗体巧妙地放置在同一反应槽内，简化试剂操作流程，同时互相之间又有物理隔断，有效避免非特异性结合。同时，通过设计该反应槽的特殊结构，使得流体经过反应槽时，能够产生类似于“波浪”的液流，促使包被抗体、标记抗体充分混合，有效地提高测试效率和灵敏度。
为实现上述的技术目的，本发明将采取如下的技术方案：
一种单通道化学发光微流控芯片，包括芯片本体，芯片本体包括光路扫描窗口、全血过滤腔、定量-反应腔以及废液腔，全血过滤腔的出液口通过微流控流道依次与定量-反应腔、废液腔连通；光路扫描窗口正对于定量-反应腔的上方设置，定量-反应腔的进液口能够分别通过清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则能够与外接气路连通；所述定量-反应腔分体设置成处于下方的定量-反应池以及能够将定量-反应池的池口封接的反应池盖板；所述定量-反应池通过垂直于流体流向所布置的两块隔板a，等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；所述反应池盖板在面向定量-反应池的板面通过垂直于流体流向所布置的隔板b，等分成两块盖板分体，每块盖板分体中均埋填有包被抗体。
作为本发明的进一步改进，所述全血过滤腔的出液口通过第一防回流装置与定量-反应腔的进液口连通，所述清洗液输送支路的出液口与显色液输送支路的出液口汇聚后，通过第二防回流装置与定量-反应池的进液口连通；所述第一防回流装置、第二防回流装置结构一致，均包括防回流结构，防回流结构位于微流控流道的上方，具有防回流流道；防回流流道能够抬升防回流结构安装位置处的微流控流道液面高度；当微流控流道接通外接气路时，在外接气路提供的气压驱动下，克服防回流流道所抬升的此处微流控流道液面高度的压力，促使防回流装置两侧的微流控流道中的流体处于流通状态。
作为本发明的进一步改进，所述定量-反应腔与废液腔之间的微流控流道上 安装有阀门装置，该阀门装置包括用于感测流体是否流经阀门装置安装位置的检测机构；在芯片本体中处于常开状态的阀门装置，在检测机构所反馈的数值表明流体流经阀门装置安装位置时，能够自动闭合，处于截流状态；处于截流状态的阀门装置能够定时打开，处于流通状态；处于流通状态的阀门装置，在外接气路提供的气流推动下，流体能够在阀门装置中流通。
作为本发明的进一步改进，所述废液腔中置放有第一吸水纸，且废液腔具有两个连通的废液腔分体，分别为废液腔分体a、废液腔分体b，其中：废液腔分体a的内壁与第一吸水纸之间能够形成气流通道，而废液腔分体b具有出气口。
作为本发明的进一步改进，所述废液腔包括废液池、能够封接废液池池口的中层盖板以及能够覆盖中层盖板的上层盖板，所述废液池在与废液腔分体a对应的池壁设置有若干废液导流件，并在紧靠着废液导流件的废液池池底安装吸纸固定柱；所述废液导流件呈弧形齿牙状，各废液导流件按照自身的弧形旋向沿着废液池的池壁依次布置，第一吸水纸通过吸纸固定柱置于废液池中，且第一吸水纸的侧边与废液导流件的尖端相触。所述中层盖板在与废液腔分体a对应的部位设置有与废液池的池口贯通的中层通孔，在与废液腔分体b对应的位置处设置有与废液池的池口连通的中层通气孔，并在中层通气孔位置处设置吸水纸嵌槽，该吸水纸嵌槽中置放有第二吸水纸；所述上层盖板在与中层通孔对应的位置处设置有能够将中层通气孔封接的通气孔封盖，且在与中层通气孔对应的位置处设置与中层通气孔连通的上层通气孔。
作为本发明的进一步改进，全血过滤腔具有能够与外接气体连接的进样部位，进样部位具有加样孔以及透气孔；所述透气孔相对于全血过滤腔底部的间距大于加样孔相对于全血过滤腔底部的间距。
作为本发明的进一步改进，进样部位具有环形凹槽和环形凸缘，该环形凸缘的外侧壁为环形凹槽的内侧壁，气源接口置于环形凹槽中；透气孔包括第一透气孔、第二透气孔；第一透气孔设于环形凸缘的端面，该环形凸缘通过一收敛过渡面与加样孔衔接，第二透气孔紧靠着环形凸缘的内壁设于收敛过渡面，且第二透气孔所处位置与第一透气孔所在位置相邻。
作为本发明的进一步改进，所述全血过滤腔包括全血滤样池以及封盖住全血滤样池的顶盖，进样部位设置于顶盖；该全血滤样池具有滤样出口，全血滤样池 的侧壁以渐缩的方式延伸至滤样出口位置处，该呈渐缩方式延伸的侧壁呈具有引流作用的齿状设置；全血滤样池中铺设有全血滤样纸，且全血滤样纸的侧边能够与相邻的全血滤样池的齿状侧壁的各齿牙内凸尖端相触。
本发明的另一技术目的是提供一种单通道化学发光微流控芯片，包括芯片本体，该芯片本体为三片式结构，包括由上到下依次层叠的上层芯片、中层芯片以及下层芯片；上层芯片、中层芯片以及下层芯片两两之间均通过定位柱、定位孔配合连接的方式实现相互间的层叠；其中：
所述下层芯片上设置有全血过滤池、定量-反应池以及废液池，全血过滤池的出液口通过微流控流道依次与定量-反应池、废液池连通，同时所述定量-反应池的进液口能够分别通过清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤池则能够与外接气路连通；
所述定量-反应池通过垂直于流体流向所布置的两块隔板a，等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；所述反应池盖板在面向定量-反应池的板面通过垂直于流体流向所布置的隔板b，等分成两块盖板分体，每块盖板分体中均埋填有包被抗体；
所述中层芯片，在与全血过滤池对应的位置处设置进样部位，在与定量-反应池对应的位置处设置能够将定量-反应池的池口封接的反应池盖板，在与废液池对应的位置处则设置有能够封接废液池池口的中层盖板，在与清洗液输送支路进液口、显色液输送支路进液口对应的位置处分别设置清洗液接口、显色液接口；
所述上层芯片，在与进样部位对应的位置处设置进样部位通孔，在与反应池盖板对应的位置处设置光路扫描窗口，而在中层盖板对应的位置处则设置上层盖板，在与清洗液接口、显色液接口对应的位置处分别设置清洗液接口通孔、显色液接口通孔；
所述全血过滤池与定量-反应池之间的微流控流道上安装第一防回流装置，而清洗液输送支路、显色液输送支路汇聚后，与定量-反应池之间的微流控流道上安装第二防回流装置；
所述第一防回流装置、第二防回流装置结构一致，均包括防回流结构，防回流结构位于微流控流道的上方，具有防回流流道；防回流流道能够抬升防回流结构安装位置处的微流控流道液面高度；当微流控流道接通外接气路时，在外接气 路提供的气压驱动下，克服防回流流道所抬升的此处微流控流道液面高度的压力，促使防回流装置两侧的微流控流道中的流体处于流通状态；
所述定量-反应池与废液池之间的微流控流道上安装阀门装置，该阀门装置包括用于感测流体是否流经阀门装置安装位置的检测机构；在芯片本体中处于常开状态的阀门装置，在检测机构所反馈的数值表明流体流经阀门装置安装位置时，能够自动闭合，处于截流状态；处于截流状态的阀门装置能够定时打开，处于流通状态；处于流通状态的阀门装置，在外接气路提供的气流推动下，流体能够在阀门装置中流通。
本发明的再一个技术目的是提供一种单通道化学发光微流控芯片的检测方法，包括以下步骤：
步骤一、向全血过滤腔的加样孔加入全血样本；
步骤二、芯片本体在孵育一段时间t1后，将芯片本体与外接气路、外接液路以及下压机构接触，使得芯片本体的清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则与外接气路连通；同时阀门装置中，下压机构的下压头与导电密封圈的内部封闭膜相触，电容探头与导电密封圈的电容探头触点相触；
步骤三、外接气路向芯片本体缓慢充气，推动全血样本在全血过滤纸的过滤下，变成血浆，进入全血过滤腔；
步骤四、在外接气路持续气压的推动下，在全血过滤腔中的血浆，沿着微流控流道继续向前流动，依次流经第一防回流装置、定量-反应腔后，流入阀门装置；此时，血浆一旦接触阀门装置的导电密封圈，致使电容探头感测到与其相触的导电密封圈的电容探头触点的电容发生变化，触动阀门装置，关闭微流控流道，在此同时，关闭外接气路，停止加压；
步骤五、将芯片本体与外接气路、外接液路以及下压机构分离；让处于定量-反应腔中的血浆与其中预埋的包被抗体、标记抗体混匀3-10分钟，进行免疫反应；
步骤六、将芯片本体与外接气路、外接液路以及下压机构接触，使得芯片本体的清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则与外接气路连通；同时阀门装置中，下压机构的下压头与导电密封圈的内部封闭 膜相触，电容探头与导电密封圈的电容探头触点相触；
步骤七、启动外接气路，外接气路提供的气压将推动芯片本体中的血浆沿着微流控流道流动，直至吹干加样孔以及微流控流道中的血浆，此时，电容探头感测到的电容值恢复至初始值，外接气路关闭；
步骤八、启动外接液路中的清洗液装置，促使清洗液依次流经清洗液输送支路上所安装的第二防回流装置、定量-反应腔后，抵达阀门装置；清洗液一旦接触阀门装置的导电密封圈，致使电容探头感测到与其相触的导电密封圈的电容探头触点的电容发生变化，触动阀门装置，关闭微流控流道，在此同时，关闭外接气路，停止加压；
步骤九、清洗液在芯片本体，尤其是定量-反应腔中混匀1-3分钟，进行清洗；
步骤十、启动外接气路，提供气压推动清洗液向前移动，吹干芯片本体中的液体，此时电容探头感测到的电容值恢复至初始值，外接气路关闭；
步骤十一、重复步骤八至步骤十3-8次；
步骤十二、启动外接液路中的显色液装置，促使显色液依次流经显色液输送支路上所安装的防回流装置、定量-反应腔后，抵达阀门装置；显色液一旦接触阀门装置的导电密封圈，致使电容探头感测到与其相触的导电密封圈的电容探头触点的电容发生变化，触动阀门装置，关闭微流控流道，在此同时，关闭外接气路，停止加压；
步骤十三、显色液在定量-反应腔中混匀3-8分钟，进行显色反应；
步骤十四、将芯片本体与外接气路、外接液路以及下压机构分离，完成检测。
根据上述的技术方案，相对于现有技术，本发明具有如下的优点：
本发明提供了一种特殊结构形式的定量-反应腔，其能够在其中同时置放包被抗体、标记抗体，同时所置放的包被抗体、标记抗体之间又存在物理隔断(隔板a、隔板b、置放于反应池盖板包被抗体与置放于定量-反应池中的标记抗体存在间隙)，因此，简化试剂操作流程，有效地避免包被抗体、标记抗体之间的非特异性结合，另外，本发明所述的定量-反应腔，通过所设置的隔板a、隔板b，可以使得流经定量-反应腔的液流呈波浪状，可以很好地将包被抗体、标记抗体混合，有效地提高提高测试效率和灵敏度。
1、本发明所述定量-反应腔的进液口前端配装防回流装置(第一防回流装置、第二防回流装置)，使得一个设备可以操作多个微流控芯片，起到多通量的效果，因为即便把液体向前流动的推动力去掉，液体也不会回流过去，因为有防回流装置，这样各个步骤之间的孵育时间时，设备可以离开微流控芯片，去操作另一个微流控芯片，起到多通量的目的。
2、本发明所述定量-反应腔的后端配装阀门装置，该阀门装置和设备紧密结合，通过有封闭膜的密封圈和设备上面的下压装置，起到随时开关阀门的目的，而且可以重复使用。同时保证大规模生产的需要，阀门装置对生产工艺的要求不太高。同时，可以及时检测流道内液体流动至密封圈的时机，准确关闭密封圈，既可排除液体前面的气泡，又可以节流液体，起到半透膜(透气不透液)装置的作用。节省半透膜装置功能模块。
3、本发明所述全血过滤腔在进样部位分别设置有加样孔、透气孔，避免因为主动式微流控而引起的全血样本堵塞的问题；将透气孔设置成高度不等的第二透气孔、第一透气孔，第二透气孔相对于全血过滤腔底部的间距大于加样孔相对于全血过滤腔底部的间距，尤其是将第一透气孔设置高度高于第二透气孔，则在加样过程中，即便加样孔、第二透气孔堵塞了，全血滤样腔还能够通过第一透气孔与气源接通，主动式微流控气路的流通；
4、本发明将全血滤样池的侧壁以渐缩的方式延伸至滤样出口位置处，可以起到引流作用，而将该呈渐缩方式延伸的侧壁呈具有引流作用的齿状设置，可以有效地避免由于生产误差导致的缝隙(全血滤样池侧壁与全血滤样纸侧边之间的缝隙)无法完全过滤全血样本的问题。
5、本发明在废液腔设置气体通道，并通过废液腔出气口将气体排出，有效地保证液流能够很好地被吸水纸吸收，防止废液回流、外溅或者液漏。
附图说明
图1是本发明所述单通道化学发光微流控芯片的分解结构示意图；
图中：1、上层芯片；2、中层芯片；3、下层芯片；4、外接气路；5、外接液路；5-1、密封垫；6-1、下压机构；6-2、导电密封圈；7-1、第二吸水纸；7-2、第一吸水纸；8、滤血纸；
图2是本发明所述下层芯片的结构示意图；
图3是图2中废液池部分(无吸水纸)的放大结构示意图；
图2、图3中：3-1、全血过滤池；3-1-1、样品导流件；3-2、定量-反应池；3-2-1、隔板a；3-3、废液池；3-3-1、废液导流件；3-3-2、吸水纸定位柱；3-4、清洗液输送支路；3-5、显色液输送支路；3-6、下层芯片外定位口；3-7、芯片固定柱；6、阀门装置；9-a、第一回流装置；9-b、第二回流装置；
图4是图2中废液池部分(装有吸水纸)的放大结构示意图；
图4中箭头走向表明废液池中的气流通道；
图5是中层芯片的一个方向(正面)的结构示意图；
图6是中层芯片另一个方向(背面)的结构示意图；
其中：2-1、加样槽；2-1-1、透气孔；2-1-2、加样孔；2-2、芯片定位孔；2-3、第一回流装置的回流孔；2-4、第二回流装置的回流孔；2-5、清洗液支路的外接接口；2-6、显色液支路的外接接口；2-7、导电密封圈的安装槽；2-8、废液池的中层通孔；2-9、出气口；2-10、反应池盖板；
图7是上层芯片正面的结构示意图；
图8是上层芯片背面的结构示意图；
图7、图8中：1-1、进样部位通孔；1-2、光路扫描窗口；1-3、废液池透气孔；1-4、清洗液外接接口的通孔；1-5、显色液外接接口的通孔；1-6、电容探头穿行孔；1-7、下压头穿行孔；
图9是三片式防回流结构的结构示意图；
图10是图9的截面图；
图11是第二种结构形式防回流结构的结构示意图；
图12是图11的截面图；
图13是第三种结构形式的防回流结构中，下层芯片的结构示意图；
图14是第三种结构形式的防回流结构中，上层芯片的结构示意图；
图9-14中：上层芯片1；中层芯片2；防回流出液口2-a；防回流连通流道2-b；防回流凸块2-c；防回流流道2-d；下层芯片3；微流控流道3-a；微流控流道隔块3-b；连接柱3-c；
图15是本发明所述阀门装置的分解结构示意图；
图16是阀门装置对应位置处的中层芯片的结构示意图；
图17是导电密封圈的立体结构示意图；
图18是图16一个方向的断面图；
图19是图16另一个方向的断面图；
图15-图19中：1、上层芯片；2、中层芯片；2-e、中层芯片流道；2-f、密封圈嵌槽；3、下层芯片；6-1、下压机构；6-2、导电密封圈；6-2-1、外层密封圈；6-2-2、内部封闭膜；6-2-3、电容探头触点；6-3、检测探头。
具体实施方式
下面将结合本实用新型实施例中的附图，对本实用新型实施例中的技术方案进行清楚、完整地描述，显然，所描述的实施例仅仅是本实用新型一部分实施例，而不是全部的实施例。以下对至少一个示例性实施例的描述实际上仅仅是说明性的，决不作为对本实用新型及其应用或使用的任何限制。基于本实用新型中的实施例，本领域普通技术人员在没有作出创造性劳动前提下所获得的所有其他实施例，都属于本实用新型保护的范围。除非另外具体说明，否则在这些实施例中阐述的部件和步骤的相对布置、表达式和数值不限制本实用新型的范围。同时，应当明白，为了便于描述，附图中所示出的各个部分的尺寸并不是按照实际的比例关系绘制的。对于相关领域普通技术人员已知的技术、方法和设备可能不作详细讨论，但在适当情况下，所述技术、方法和设备应当被视为授权说明书的一部分。在这里示出和讨论的所有示例中，任何具体值应被解释为仅仅是示例性的，而不是作为限制。因此，示例性实施例的其它示例可以具有不同的值。
为了便于描述，在这里可以使用空间相对术语，如“在……之上”、“在……上方”、“在……上表面”、“上面的”等，用来描述如在图中所示的一个器件或特征与其他器件或特征的空间位置关系。应当理解的是，空间相对术语旨在包含除了器件在图中所描述的方位之外的在使用或操作中的不同方位。例如，如果附图中的器件被倒置，则描述为“在其他器件或构造上方”或“在其他器件或构造之上”的器件之后将被定位为“在其他器件或构造下方”或“在其他器件或构造之下”。因而，示例性术语“在……上方”可以包括“在……上方”和“在……下方”两种方位。该器件也可以其他不同方式定位(旋转90度或处于其他方位)。
本发明公开了一种单通道化学发光微流控芯片，包括芯片本体，芯片本体包括光路扫描窗口、全血过滤腔、定量-反应腔以及废液腔，全血过滤腔的出液口 通过微流控流道依次与定量-反应腔、废液腔连通；光路扫描窗口正对于定量-反应腔的上方设置，定量-反应腔的进液口能够分别通过清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则能够与外接气路连通；所述定量-反应腔分体设置成处于下方的定量-反应池以及能够将定量-反应池的池口封接的反应池盖板；所述定量-反应池通过垂直于流体流向所布置的两块隔板a，等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；所述反应池盖板在面向定量-反应池的板面通过垂直于流体流向所布置的隔板b，等分成两块盖板分体，每块盖板分体中均埋填有包被抗体。
所述全血过滤腔的出液口通过第一防回流装置与定量-反应腔的进液口连通，所述清洗液输送支路的出液口与显色液输送支路的出液口汇聚后，通过第二防回流装置与定量-反应池的进液口连通；所述第一防回流装置、第二防回流装置结构一致，均包括防回流结构，防回流结构位于微流控流道的上方，具有防回流流道；防回流流道能够抬升防回流结构安装位置处的微流控流道液面高度；当微流控流道接通外接气路时，在外接气路提供的气压驱动下，克服防回流流道所抬升的此处微流控流道液面高度的压力，促使防回流装置两侧的微流控流道中的流体处于流通状态。
所述定量-反应腔与废液腔之间的微流控流道上安装有阀门装置，该阀门装置包括用于感测流体是否流经阀门装置安装位置的检测机构；在芯片本体中处于常开状态的阀门装置，在检测机构所反馈的数值表明流体流经阀门装置安装位置时，能够自动闭合，处于截流状态；处于截流状态的阀门装置能够定时打开，处于流通状态；处于流通状态的阀门装置，在外接气路提供的气流推动下，流体能够在阀门装置中流通。
所述废液腔中置放有第一吸水纸，且废液腔具有两个连通的废液腔分体，分别为废液腔分体a、废液腔分体b，其中：废液腔分体a的内壁与第一吸水纸之间能够形成气流通道，而废液腔分体b具有出气口。
所述废液腔包括废液池、能够封接废液池池口的中层盖板以及能够覆盖中层盖板的上层盖板，所述废液池在与废液腔分体a对应的池壁设置有若干废液导流件，并在紧靠着废液导流件的废液池池底安装吸纸固定柱；所述废液导流件呈弧形齿牙状，各废液导流件按照自身的弧形旋向沿着废液池的池壁依次布置，第一 吸水纸通过吸纸固定柱置于废液池中，且第一吸水纸的侧边与废液导流件的尖端相触。所述中层盖板在与废液腔分体a对应的部位设置有与废液池的池口贯通的中层通孔，在与废液腔分体b对应的位置处设置有与废液池的池口连通的中层通气孔，并在中层通气孔位置处设置吸水纸嵌槽，该吸水纸嵌槽中置放有第一吸水纸；所述上层盖板在与中层通孔对应的位置处设置有能够将中层通气孔封接的通气孔封盖，且在与中层通气孔对应的位置处设置与中层通气孔连通的上层通气孔。
全血过滤腔具有能够与外接气体连接的进样部位，进样部位具有加样孔以及透气孔；所述透气孔相对于全血过滤腔底部的间距大于加样孔相对于全血过滤腔底部的间距。
进样部位具有环形凹槽和环形凸缘，该环形凸缘的外侧壁为环形凹槽的内侧壁，气源接口置于环形凹槽中；透气孔包括第一透气孔、第二透气孔；第一透气孔设于环形凸缘的端面，该环形凸缘通过一收敛过渡面与加样孔衔接，第二透气孔紧靠着环形凸缘的内壁设于收敛过渡面，且第二透气孔所处位置与第一透气孔所在位置相邻。
所述全血过滤腔包括全血滤样池以及封盖住全血滤样池的顶盖，进样部位设置于顶盖；该全血滤样池具有滤样出口，全血滤样池的侧壁以渐缩的方式延伸至滤样出口位置处，该呈渐缩方式延伸的侧壁呈具有引流作用的齿状设置；全血滤样池中铺设有全血滤样纸，且全血滤样纸的侧边能够与相邻的全血滤样池的齿状侧壁的各齿牙内凸尖端相触。
附图详细地公开了本发明所述的一个具体实施例，以下将结合附图详细地说明本发明的技术方案。
如图1至19所示，本发明所述的单通道化学发光微流控芯片，包括芯片本体，该芯片本体为三片式结构，包括由上到下依次层叠的上层芯片、中层芯片以及下层芯片；上层芯片、中层芯片以及下层芯片两两之间均通过定位柱、定位孔配合连接的方式实现相互间的层叠；其中：
所述下层芯片，如图2、图3所示，设置有全血过滤池、定量-反应池以及废液池，全血过滤池的出液口通过微流控流道依次与定量-反应池、废液池连通，同时所述定量-反应池的进液口能够分别通过清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤池则能够与外接气路连通。
所述全血滤样池具有滤样出口，全血滤样池的侧壁以渐缩的方式延伸至滤样出口位置处，该呈渐缩方式延伸的侧壁呈具有引流作用的齿状设置；全血滤样池中铺设有全血滤样纸，且全血滤样纸的侧边能够与相邻的全血滤样池的齿状侧壁的各齿牙内凸尖端相触。全血滤样纸通过进样部位下端所设置的滤样纸压点压紧在全血滤样池的槽底。
所述定量-反应池，通过垂直于流体流向所布置的两块隔板a，等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；所述反应池盖板在面向定量-反应池的板面通过垂直于流体流向所布置的隔板b，等分成两块盖板分体，每块盖板分体中均埋填有包被抗体。
所述废液池，如图4所示，具有连通的废液池分体a、废液池分体b，废液池分体a的池壁设置有若干废液导流件，并在紧靠着废液导流件的池底安装吸纸固定柱；所述废液导流件呈弧形齿牙状，各废液导流件按照自身的弧形旋向沿着废液池分体a的池壁依次布置，第一吸水纸通过吸纸固定柱置于废液池中，且第一吸水纸的侧边与废液导流件的尖端相触。
所述中层芯片，如图5、图6所示，在与全血过滤池对应的位置处设置进样部位，在与定量-反应池对应的位置处设置能够将定量-反应池的池口封接的反应池盖板，在与废液池对应的位置处则设置有能够封接废液池池口的中层盖板，在与清洗液输送支路进液口、显色液输送支路进液口对应的位置处分别设置清洗液接口、显色液接口；所述中层盖板在与废液池分体a对应的部位设置有贯通的中层通孔，在与废液池分体b对应的位置处设置有中层通气孔，并在中层通气孔位置处设置吸水纸嵌槽，该吸水纸嵌槽中置放有第二吸水纸。
进样部位具有环形凹槽和环形凸缘，该环形凸缘的外侧壁为环形凹槽的内侧槽壁，环形凸缘设置于中层芯片的上表面，环形凹槽的外侧槽壁设置于上层芯片，而中层芯片在环形凹槽外侧槽壁与环形凸缘的外侧壁之间设置有环形凹槽的槽底，气源接口置于环形凹槽中；透气孔包括第一透气孔、第二透气孔；第一透气孔设于环形凸缘的端面，该环形凸缘通过锥形过渡面与加样孔衔接，第二透气孔紧靠着环形凸缘的内壁设于锥形过渡面。
所述上层芯片，如图7、图8所示，在与进样部位对应的位置处设置进样部位通孔，在与反应池盖板对应的位置处设置光路扫描窗口，而在中层盖板对应的 位置处则设置上层盖板，在与清洗液接口、显色液接口对应的位置处分别设置清洗液接口通孔、显色液接口通孔；所述上层盖板在与中层通孔对应的位置处设置有能够将中层通气孔封接的通气孔封盖，且在与中层通气孔对应的位置处设置与中层通气孔连通的上层通气孔。
如图2所示，所述全血过滤池与定量-反应池之间的微流控流道上安装第一防回流装置，而清洗液输送支路、显色液输送支路汇聚后，与定量-反应池之间的微流控流道上安装第二防回流装置。
所述第一防回流装置、第二防回流装置结构一致，如图9-14所示，均包括防回流结构，防回流结构位于微流控流道的上方，具有防回流流道；防回流流道能够抬升防回流结构安装位置处的微流控流道液面高度；当微流控流道接通外接气路时，在外接气路提供的气压驱动下，克服防回流流道所抬升的此处微流控流道液面高度的压力，促使防回流装置两侧的微流控流道中的流体处于流通状态。
所述防回流流道的延伸方向与防回流结构安装位置处微流控流道的延伸方向相垂直；防回流结构安装位置处的微流控流道通过微流控流道隔块分隔成两段，分别为微流控进液流道、微流控出液流道；防回流流道的上端封闭，下端则跨接在微流控流道隔块上方，并分别与微流控进液流道、微流控出液流道连通。
所述防回流流道内具有防回流凸块，且防回流凸块与防回流流道之间形成有两条防回流连通流道；所述的两条防回流连通流道，分别与微流控进液流道、微流控出液流道连通；防回流凸块的下端与防回流流道的下端齐平，而防回流凸块的上端则低于防回流流道的上端设置。
所述防回流凸块在两条防回流连通流道之间的截面成梯形状设置。
所述微流控流道隔块为楔形块，而防回流流道为形状与微流控流道隔块形状相似的楔形槽。
防回流结构安装位置处的微流控流道，在微流控流道隔块的两侧对称地布置有楔形连接柱，防回流流道具有与楔形连接柱配合连接的楔形连接槽。
如图2所示，所述定量-反应池与废液池之间的微流控流道上安装阀门装置，该阀门装置，如图15-19所示，包括用于感测流体是否流经阀门装置安装位置的检测机构；在芯片本体中处于常开状态的阀门装置，在检测机构所反馈的数值表明流体流经阀门装置安装位置时，能够自动闭合，处于截流状态；处于截流状态 的阀门装置能够定时打开，处于流通状态；处于流通状态的阀门装置，在外接气路提供的气流推动下，流体能够在阀门装置中流通。
所述阀门装置为具有防回流功能的防回流微阀；包括下压机构、检测机构、密封件以及微阀防回流结构；其中：
所述的下压机构，包括下压头，该下压头位于内部封闭膜的上方。
所述检测机构，包括检测探头，该检测探头与下压头联动连接，并位于探头触点的上方，同时检测探头与探头触点之间的间距小于下压头与内部封闭膜之间的间距。
所述的密封件为导电密封圈，包括外层密封圈、内部封闭膜以及电容探头触点；内部封闭膜处于外层密封圈的内圈，而电容探头触点则与外层密封圈连接。
所述微阀防回流结构，与防回流微阀安装位置处的微流控流道分体设置，包括贯通的微阀防回流流道；
所述防回流微阀安装位置处的微流控流道通过微流控流道横隔分成微阀进液段和微阀出液段；微阀进液段具有出液口，微阀出液段具有进液口；
微阀防回流流道的下端置于微流控流道横隔的上方，且微阀防回流流道能够分别与微阀进液段的出液口、微阀出液段的进液口对应连通；
本发明中，微阀防回流流道的内腔具有微阀防回流凸块；微阀防回流凸块的外圈与微阀防回流流道的内腔之间形成用于安装外层密封圈的密封圈嵌槽；微阀防回流凸块具有两个贯通流道，分别为微阀防回流进液流道、微阀防回流出液流道；微阀防回流进液流道的下端与微阀进液段的出液口连通，而微阀防回流出液流道的下端则与微阀出液段的进液口连通；外层密封圈嵌装在密封圈嵌槽中，内部封闭膜能够同时覆盖住微阀防回流进液流道的上端、微阀防回流出液流道的上端；
当本发明不配装防回流结构时，所述的内部封闭膜能够同时覆盖住微阀进液段的出液口、微阀出液段的进液口，并通过外层密封圈密封。
检测机构的检测探头随着下压机构的下压动作，与电容探头触点相触；随时检测液体是否流到阀门内。此时，防回流微阀处于打开状态。事实上，在下压机构的下压头与内部封闭膜之间存在间距的情况下，由于内部封闭膜本身具有的形变功能以及安装位置处具有的形变空间，致使在气流的带动下，流体能够从微阀 进液段中，克服内部封闭膜形变后，进入微阀出液段。
当接通气路时，在气路提供的气压驱动下，液体向前移动，微阀进液段中的流体经由微阀防回流流道后，流至微阀出液段，检测机构中的电容探头，检测到液体流过来，再次触发下压机构1，在下压驱动的带动下，内部封闭膜封住微阀防回流流道的的上端，此时，防回流微阀处于关闭状态；
检测机构包括电容探头，电容探头在下压机构的下压驱动的带动下，能够与电容探头触点电性连接。
具体地：当下压机构的下压头带着检测探头面向导电密封圈下行时，在检测探头与探头触点相触时，下压机构停止下行，此时，下压头与内部封闭膜之间存在间距；当检测探头感测的数据出现变化，表明存在流体流经阀门装置安装位置处的微流控流道时，触发下压机构动作，推动下压头继续下行，直至下压头压住内部封闭膜，以同时封住微阀进液段的出液口、微阀出液段的进液口，此时阀门装置处于截流状态；当下压机构的下压头与内部封闭膜之间存在间隙时，阀门装置处于打开状态，内部封闭膜所具有的形变特征，确保流体能够从微阀进液段的出液口流出，从微阀出液段的进液口流入。
附图中，所述阀门装置为三片式结构，包括从上到下依次分布的上层芯片、中层芯片以及下次芯片；上层芯片、中层芯片以及下次芯片两两相互卡合连接；微阀防回流结构设置于中层芯片，微流控流道布置于下层芯片，嵌槽对应于微阀防回流结构设置于中层芯片的上表面，密封件通过上层芯片压设在嵌槽中，且上层芯片对应于内部密封圈、检测触点的位置对应地分别开设压头穿行孔、探头穿行孔；而下压机构位于上层芯片的上方，下压机构下压，带动下压头通过压头穿行孔，压住内部封闭膜，以封住微阀防回流流道的上端，此时，防回流微阀处于关闭状态；电容探头随着下压机构1的下压而朝向上层芯片移动，直至穿过上层芯片上的探头穿行孔后，与置于中层芯片嵌槽中的导电密封圈的探头触点相触，实现电性连接。
本发明创造性地设计了阀门装置，使其和微流控液体控制防回流装置融合在一起，节省微流控芯片空间。微流控芯片的位置空间十分有限，本发明能有效的节省微流控流道空间，可以使设计流道时更加有效。避免各个流道之间距离过近导致的液体流道封闭不严，穿流道的问题。
本发明利用导电橡胶材质的导电性，结合设备的电容检测探头，观察流道内液体流动到阀门的准确时机，及时关闭阀门。节省半透膜装置。
基于上述的单通道微流控芯片，本发明提供了如下的一种检测方法，具体包括如下步骤：
步骤1、试剂芯片插入仪器内，读取上面信息；
步骤2、移液器加样，100ul的全血加入加样孔内；
步骤3、芯片进入仪器内部，孵育2分钟；
步骤4、仪器与芯片的接触装置结合，含有气路装置，两个液路装置，带有导电橡胶探头的阀门压点；
步骤5、气路装置缓慢充气，推动全血样本往前移动，在此过程中，红细胞被滤血纸截留，但是由于气体压力，红细胞堵塞，加样孔部分可能会通气不畅，此时气体由透气口进入向前推动样本前进；
步骤6、样本通过滤血纸后，变为血浆，进入流道内，再由压力推动通过防回流装置，然后样本会通过液体流道，但是由于液体流道终端有液路装置紧压芯片，液体无法进入此通道，而且液体流道和样本流动方向也不同；
步骤7、样本继续向前进入反应定量孔内；
步骤8、样本继续向前，进入阀门装置内，样本一旦接触阀门内导电橡胶，电容变化会触动阀门开关，关闭流道阀门，此时在阀门后面又有一个防回流装置，避免阀门关闭不及时，液体很快吹走的问题。同时关闭气路开关，停止加压；
步骤9、仪器和芯片接触装置分离，混匀3-10分钟，进行免疫反应；
步骤10、仪器和芯片接触装置结合，气体推动样本进行向前移动，吹干加样孔和流道样本，此时导电橡胶探头数据变回原来的值，关闭气路装置；
步骤11、清洗液路装置启动，进入流道内，再由压力推动通过防回流装置，然后清洗液会通过样本流道，但是由于样本流道终端有气路装置紧压芯片，液体无法进入此通道，而且液体流道和样本流动方向也不同。同时，另一路液路通道由于液体流道终端有液路装置紧压芯片，液体也无法进入此通道，而且液体流道和样本流动方向也不同；
步骤12、清洗液继续向前进入反应定量孔内；
步骤13、清洗液继续向前，进入阀门装置内，清洗液一旦接触阀门内导电 橡胶，电容变化会触动阀门开关，关闭流道阀门，此时在阀门后面又有一个防回流装置，避免阀门关闭不及时，液体很快吹走的问题。同时关闭气路开关，停止加压；
步骤14、混匀1-3分钟，进行清洗；
步骤15、气体推动清洗液进行向前移动，吹干加样孔和流道液体，此时导电橡胶探头数据变回原来的值，关闭气路装置；
步骤16、重复11到15步骤3-8次；
步骤17、显色液液路装置启动，进入流道内，再由压力推动通过防回流装置，然后清洗液会通过样本流道，但是由于样本流道终端有气路装置紧压芯片，液体无法进入此通道，而且液体流道和样本流动方向也不同。同时，另一路液路通道由于液体流道终端有液路装置紧压芯片，液体也无法进入此通道，而且液体流道和样本流动方向也不同；
步骤18、显色液继续向前进入反应定量孔内；
步骤19、清洗液继续向前，进入阀门装置内，清洗液一旦接触阀门内导电橡胶，电容变化会触动阀门开关，关闭流道阀门，此时在阀门后面又有一个防回流装置，避免阀门关闭不及时，液体很快吹走的问题。同时关闭气路开关，停止加压；
步骤20、混匀3-8分钟，进行显色反应；
步骤21、仪器和芯片接触装置分离；
步骤22、仪器读取数据；
步骤23、仪器退出芯片，完成检测。

Claims (10)

  1. 一种单通道化学发光微流控芯片，包括芯片本体，芯片本体包括光路扫描窗口、全血过滤腔、定量-反应腔以及废液腔，全血过滤腔的出液口通过微流控流道依次与定量-反应腔、废液腔连通；光路扫描窗口正对于定量-反应腔的上方设置，定量-反应腔的进液口能够分别通过清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则能够与外接气路连通；其特征在于，所述定量-反应腔分体设置成处于下方的定量-反应池以及能够将定量-反应池的池口封接的反应池盖板；所述定量-反应池通过垂直于流体流向所布置的两块隔板a，等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；所述反应池盖板在面向定量-反应池的板面通过垂直于流体流向所布置的隔板b，等分成两块盖板分体，每块盖板分体中均埋填有包被抗体。
  2. 根据权利要求1所述的单通道化学发光微流控芯片，其特征在于，所述全血过滤腔的出液口通过第一防回流装置与定量-反应腔的进液口连通，所述清洗液输送支路的出液口与显色液输送支路的出液口汇聚后，通过第二防回流装置与定量-反应池的进液口连通；所述第一防回流装置、第二防回流装置结构一致，均包括防回流结构，防回流结构位于微流控流道的上方，具有防回流流道；防回流流道能够抬升防回流结构安装位置处的微流控流道液面高度；当微流控流道接通外接气路时，在外接气路提供的气压驱动下，克服防回流流道所抬升的此处微流控流道液面高度的压力，促使防回流装置两侧的微流控流道中的流体处于流通状态。
  3. 根据权利要求1所述的单通道化学发光微流控芯片，其特征在于，所述定量-反应腔与废液腔之间的微流控流道上安装有阀门装置，该阀门装置包括用于感测流体是否流经阀门装置安装位置的检测机构；在芯片本体中处于常开状态的阀门装置，在检测机构所反馈的数值表明流体流经阀门装置安装位置时，能够自动闭合，处于截流状态；处于截流状态的阀门装置能够定时打开，处于流通状态；处于流通状态的阀门装置，在外接气路提供的气流推动下，流体能够在阀门装置中流通。
  4. 根据权利要求1所述的单通道化学发光微流控芯片，其特征在于，所述废液腔中置放有第一吸水纸，且废液腔具有两个连通的废液腔分体，分别为废液 腔分体a、废液腔分体b，其中：废液腔分体a的内壁与第一吸水纸之间能够形成气流通道，而废液腔分体b具有出气口。
  5. 根据权利要求4所述的单通道化学发光微流控芯片，其特征在于，所述废液腔包括废液池、能够封接废液池池口的中层盖板以及能够覆盖中层盖板的上层盖板，所述废液池在与废液腔分体a对应的池壁设置有若干废液导流件，并在紧靠着废液导流件的废液池池底安装吸纸固定柱；所述废液导流件呈弧形齿牙状，各废液导流件按照自身的弧形旋向沿着废液池的池壁依次布置，第一吸水纸通过吸纸固定柱置于废液池中，且第一吸水纸的侧边与废液导流件的尖端相触；所述中层盖板在与废液腔分体a对应的部位设置有与废液池的池口贯通的中层通孔，在与废液腔分体b对应的位置处设置有与废液池的池口连通的中层通气孔，并在中层通气孔位置处设置吸水纸嵌槽，该吸水纸嵌槽中置放有第二吸水纸；所述上层盖板在与中层通孔对应的位置处设置有能够将中层通气孔封接的通气孔封盖，且在与中层通气孔对应的位置处设置与中层通气孔连通的上层通气孔。
  6. 根据权利要求1所述的单通道化学发光微流控芯片，其特征在于，全血过滤腔具有能够与外接气体连接的进样部位，进样部位具有加样孔以及透气孔；所述透气孔相对于全血过滤腔底部的间距大于加样孔相对于全血过滤腔底部的间距。
  7. 根据权利要求6所述的单通道化学发光微流控芯片，其特征在于，进样部位具有环形凹槽和环形凸缘，该环形凸缘的外侧壁为环形凹槽的内侧壁，气源接口置于环形凹槽中；透气孔包括第一透气孔、第二透气孔；第一透气孔设于环形凸缘的端面，该环形凸缘通过一收敛过渡面与加样孔衔接，第二透气孔紧靠着环形凸缘的内壁设于收敛过渡面，且第二透气孔所处位置与第一透气孔所在位置相邻。
  8. 根据权利要求6所述的单通道化学发光微流控芯片，其特征在于，所述全血过滤腔包括全血滤样池以及封盖住全血滤样池的顶盖，进样部位设置于顶盖；该全血滤样池具有滤样出口，全血滤样池的侧壁以渐缩的方式延伸至滤样出口位置处，该呈渐缩方式延伸的侧壁呈具有引流作用的齿状设置；全血滤样池中铺设有全血滤样纸，且全血滤样纸的侧边能够与相邻的全血滤样池的齿 状侧壁的各齿牙内凸尖端相触。
  9. 一种单通道化学发光微流控芯片，包括芯片本体，其特征在于，该芯片本体为三片式结构，包括由上到下依次层叠的上层芯片、中层芯片以及下层芯片；上层芯片、中层芯片以及下层芯片两两之间均通过定位柱、定位孔配合连接的方式实现相互间的层叠；其中：
    所述下层芯片上设置有全血过滤池、定量-反应池以及废液池，全血过滤池的出液口通过微流控流道依次与定量-反应池、废液池连通，同时所述定量-反应池的进液口能够分别通过清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤池则能够与外接气路连通；
    所述定量-反应池通过垂直于流体流向所布置的两块隔板a，等分成三个隔腔，其中处于中部的隔腔中置放有标记抗体，而处于两侧的隔腔则置放包被抗体；所述反应池盖板在面向定量-反应池的板面通过垂直于流体流向所布置的隔板b，等分成两块盖板分体，每块盖板分体中均埋填有包被抗体；
    所述中层芯片，在与全血过滤池对应的位置处设置进样部位，在与定量-反应池对应的位置处设置能够将定量-反应池的池口封接的反应池盖板，在与废液池对应的位置处则设置有能够封接废液池池口的中层盖板，在与清洗液输送支路进液口、显色液输送支路进液口对应的位置处分别设置清洗液接口、显色液接口；
    所述上层芯片，在与进样部位对应的位置处设置进样部位通孔，在与反应池盖板对应的位置处设置光路扫描窗口，而在中层盖板对应的位置处则设置上层盖板，在与清洗液接口、显色液接口对应的位置处分别设置清洗液接口通孔、显色液接口通孔；
    所述全血过滤池与定量-反应池之间的微流控流道上安装第一防回流装置，而清洗液输送支路、显色液输送支路汇聚后，与定量-反应池之间的微流控流道上安装第二防回流装置；
    所述第一防回流装置、第二防回流装置结构一致，均包括防回流结构，防回流结构位于微流控流道的上方，具有防回流流道；防回流流道能够抬升防回流结构安装位置处的微流控流道液面高度；当微流控流道接通外接气路时，在外接气路提供的气压驱动下，克服防回流流道所抬升的此处微流控流道液 面高度的压力，促使防回流装置两侧的微流控流道中的流体处于流通状态；
    所述定量-反应池与废液池之间的微流控流道上安装阀门装置，该阀门装置包括用于感测流体是否流经阀门装置安装位置的检测机构；在芯片本体中处于常开状态的阀门装置，在检测机构所反馈的数值表明流体流经阀门装置安装位置时，能够自动闭合，处于截流状态；处于截流状态的阀门装置能够定时打开，处于流通状态；处于流通状态的阀门装置，在外接气路提供的气流推动下，流体能够在阀门装置中流通。
  10. 一种单通道化学发光微流控芯片的检测方法，其特征在于，包括以下步骤：
    步骤一、向全血过滤腔的加样孔加入全血样本；
    步骤二、芯片本体在孵育一段时间t1后，将芯片本体与外接气路、外接液路以及下压机构接触，使得芯片本体的清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则与外接气路连通；同时阀门装置中，下压机构的下压头与导电密封圈的内部封闭膜相触，电容探头与导电密封圈的电容探头触点相触；
    步骤三、外接气路向芯片本体缓慢充气，推动全血样本在全血过滤纸的过滤下，变成血浆，进入全血过滤腔；
    步骤四、在外接气路持续气压的推动下，在全血过滤腔中的血浆，沿着微流控流道继续向前流动，依次流经第一防回流装置、定量-反应腔后，流入阀门装置；此时，血浆一旦接触阀门装置的导电密封圈，致使电容探头感测到与其相触的导电密封圈的电容探头触点的电容发生变化，触动阀门装置，关闭微流控流道，在此同时，关闭外接气路，停止加压；
    步骤五、将芯片本体与外接气路、外接液路以及下压机构分离；让处于定量-反应腔中的血浆与其中预埋的包被抗体、标记抗体混匀3-10分钟，进行免疫反应；
    步骤六、将芯片本体与外接气路、外接液路以及下压机构接触，使得芯片本体的清洗液输送支路、显色液输送支路与对应的外接液路连通，而全血过滤腔则与外接气路连通；同时阀门装置中，下压机构的下压头与导电密封圈的内部封闭膜相触，电容探头与导电密封圈的电容探头触点相触；
    步骤七、启动外接气路，外接气路提供的气压将推动芯片本体中的血浆沿着微流控流道流动，直至吹干加样孔以及微流控流道中的血浆，此时，电容探头感测到的电容值恢复至初始值，外接气路关闭；
    步骤八、启动外接液路中的清洗液装置，促使清洗液依次流经清洗液输送支路上所安装的第二防回流装置、定量-反应腔后，抵达阀门装置；清洗液一旦接触阀门装置的导电密封圈，致使电容探头感测到与其相触的导电密封圈的电容探头触点的电容发生变化，触动阀门装置，关闭微流控流道，在此同时，关闭外接气路，停止加压；
    步骤九、清洗液在芯片本体，尤其是定量-反应腔中混匀1-3分钟，进行清洗；
    步骤十、启动外接气路，提供气压推动清洗液向前移动，吹干芯片本体中的液体，此时电容探头感测到的电容值恢复至初始值，外接气路关闭；
    步骤十一、重复步骤八至步骤十3-8次；
    步骤十二、启动外接液路中的显色液装置，促使显色液依次流经显色液输送支路上所安装的防回流装置、定量-反应腔后，抵达阀门装置；显色液一旦接触阀门装置的导电密封圈，致使电容探头感测到与其相触的导电密封圈的电容探头触点的电容发生变化，触动阀门装置，关闭微流控流道，在此同时，关闭外接气路，停止加压；
    步骤十三、显色液在定量-反应腔中混匀3-8分钟，进行显色反应；
    步骤十四、将芯片本体与外接气路、外接液路以及下压机构分离，完成检测。
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