WO2018218738A1 - 一种毛细管化学发光检测装置及其检测方法 - Google Patents

一种毛细管化学发光检测装置及其检测方法 Download PDF

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Publication number
WO2018218738A1
WO2018218738A1 PCT/CN2017/092446 CN2017092446W WO2018218738A1 WO 2018218738 A1 WO2018218738 A1 WO 2018218738A1 CN 2017092446 W CN2017092446 W CN 2017092446W WO 2018218738 A1 WO2018218738 A1 WO 2018218738A1
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liquid
capillary
reagent
cup
capillary body
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PCT/CN2017/092446
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English (en)
French (fr)
Inventor
冉鹏
韩子华
唐云轩
王鹏
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成都普利泰生物科技有限公司
成都斯马特科技有限公司
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Priority to US16/319,027 priority Critical patent/US11105798B2/en
Priority to EP17911892.2A priority patent/EP3637104A4/en
Publication of WO2018218738A1 publication Critical patent/WO2018218738A1/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
    • G01N33/5302—Apparatus specially adapted for immunological test procedures
    • 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
    • B01L3/52—Containers specially adapted for storing or dispensing a reagent
    • B01L3/523—Containers specially adapted for storing or dispensing a reagent with means for closing or opening
    • 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
    • B01L3/52—Containers specially adapted for storing or dispensing a reagent
    • B01L3/527—Containers specially adapted for storing or dispensing a reagent for a plurality of reagents
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L9/00—Supporting devices; Holding devices
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/11—Filling or emptying of cuvettes
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/76—Chemiluminescence; Bioluminescence
    • 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
    • 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
    • B01L2200/16—Reagents, handling or storing thereof
    • 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
    • B01L2300/041—Connecting closures to device or container
    • B01L2300/044—Connecting closures to device or container pierceable, e.g. films, membranes
    • 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
    • B01L2300/0672—Integrated piercing tool
    • 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
    • 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
    • B01L2300/0832—Geometry, shape and general structure cylindrical, tube shaped
    • B01L2300/0838—Capillaries
    • 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
    • B01L2300/0861—Configuration of multiple channels and/or chambers in a single devices
    • B01L2300/0867—Multiple inlets and one sample wells, e.g. mixing, dilution
    • 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
    • B01L2300/12—Specific details about materials
    • 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
    • B01L2300/12—Specific details about materials
    • B01L2300/123—Flexible; Elastomeric
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/04—Moving fluids with specific forces or mechanical means
    • B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
    • B01L2400/0406—Moving fluids with specific forces or mechanical means specific forces capillary forces
    • B—PERFORMING OPERATIONS; TRANSPORTING
    • B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
    • B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
    • B01L2400/00—Moving or stopping fluids
    • B01L2400/06—Valves, specific forms thereof
    • B01L2400/0677—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers
    • B01L2400/0683—Valves, specific forms thereof phase change valves; Meltable, freezing, dissolvable plugs; Destructible barriers mechanically breaking a wall or membrane within a channel or chamber
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N2021/0325—Cells for testing reactions, e.g. containing reagents
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/03—Cuvette constructions
    • G01N2021/0346—Capillary cells; Microcells
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01—Arrangements or apparatus for facilitating the optical investigation
    • G01N21/11—Filling or emptying of cuvettes
    • G01N2021/115—Washing; Purging

Definitions

  • the invention belongs to the field of chemiluminescence detection, and in particular relates to a capillary chemiluminescence detecting device and a detecting method thereof.
  • the disposable spotting capillary has the capillary phenomenon of automatically sucking the liquid, which can realize automatic liquid addition, but it is only effective when it is used for the first time. When used once, the same capillary again uses the capillary phenomenon to add liquid, which will cause the liquid to be used later. Source pollution.
  • a capillary is used as a disposable reaction container, a multi-step reaction such as an immunoassay cannot be performed. During the immunoassay, two antibodies or antigens are used to bind the antigen or antibody, and a lotion is used. General reagents such as substrates are used in an auxiliary experiment. If it is used as a fixed reaction vessel, it is necessary to add a cleaning device and a cleaning process.
  • the technical problem to be solved by the present invention is that in the prior art, when a capillary is used as a disposable reaction container, a multi-step reaction such as an immunoassay cannot be performed, and a capillary can be used as a container for an immune reaction, which has high cost and work stability.
  • the problem of insufficient liquid source pollution is to provide a capillary chemiluminescence detecting device and a detecting method thereof, which can not only effectively utilize the capillary as a disposable reaction container, reduce the amount of reagents, and can effectively solve the pollution problem of the reagent source.
  • the small volume of the capillary itself can be utilized to control the reaction volume and reduce the need for the accuracy of the sample. The invention not only reduces the cost of the instrument, but also improves the reliability of the operation.
  • a capillary chemiluminescence detecting device comprising a liquid adding device for adding a reagent to a capillary body; the liquid adding device comprising a funnel cup having a bottom end communicating with a top end of the capillary body, and a mounting frame having more than one trough body, the setting a liquid reagent cup for containing a reagent in each tank; the liquid reagent cup is composed of a cup having an opening, a reagent disposed in the cup, and a sealing film disposed at an opening of the cup; the tank body The bottom end is connected to the top of the funnel cup. The inner bottom end of the tank body is further provided with a piercing device for piercing the sealing film when the liquid reagent cup is pressed to cause the reagent in the liquid reagent cup to flow into the funnel cup.
  • the liquid reagent cup opening of the invention is placed downside down in the liquid adding device, and the corresponding liquid adding device below the liquid reagent cup is designed with a puncture device for facilitating the breakage of the sealing film.
  • a puncture device for facilitating the breakage of the sealing film.
  • the liquid When the gravity of the liquid is greater than the capillary force, the liquid will flow out from the lower mouth of the capillary, and the liquid will decrease until the gravity and the capillary force are equal, and the flow stops, and the steady state is maintained in the capillary.
  • the capillary liquid release mechanism on the manual or supporting instrument is pressed down to blow out the gas with a large pressure, and the liquid in the capillary is blown out from the lower mouth and falls into the bottom waste liquid collector.
  • the blowing process lasts for 2s-10s, which ensures that all the liquid is blown out.
  • the capillary has a capillary phenomenon, which is a phenomenon in a capillary that is small enough to be compared with the radius of curvature of the liquid meniscus.
  • the entire liquid surface in the capillary will become curved, and the liquid-solid molecules The interaction can be extended to the entire liquid, and the surface of the liquid resembles a tensioned rubber film. If the liquid surface is curved, it has a tendency to flatten, so the liquid is applied to the liquid below, and the liquid is facing downward. The liquid is subjected to pressure. If the liquid level of the immersion liquid in the capillary is concave, it applies a pulling force to the liquid below, causing the liquid to rise along the wall of the tube. When the upward pulling force is equal to the gravity of the liquid column in the tube, the liquid in the tube stops rising. ,Achieve balance.
  • the present invention integrates the disposable reaction vessel capillary body with a liquid addition device with a plurality of liquid reagent cups, and allows the liquid reagent to be individually packaged into the liquid reagent cup in a single reaction requirement, and only needs to follow
  • the experimental procedure sequentially squeezes the liquid reagent cup and releases the liquid into the capillary body for reaction, thereby achieving the purpose of performing a chemiluminescence reaction on a separate capillary.
  • the above method can solve the problem of contamination of the reagent source when the disposable capillary is added to a plurality of reagents, and can also omit the measurement operation of the immunoreactive reagent in the immune reaction, reduce the labor intensity of the immunoreactive reagent, and avoid the amount.
  • the human error in the process is taken to improve the stability of the detection.
  • the invention also utilizes the small volume of the capillary itself to realize the control of the reaction volume, not only can achieve the effect of reducing the amount of reagents in the detection by using the capillary structure in the prior art, but also through the optimization and structure of the invention.
  • the combination of the principles of the capillary phenomenon can also effectively reduce the need for the accuracy of the sample loading. Therefore, the present invention reduces the cost of the instrument and improves the reliability of the operation as compared with the prior art.
  • the capillary body is coated with an antibody or an antigen
  • the number of the tanks on the mounting frame is set to three or more, and one reagent is used in each liquid reagent cup, and the reagent in the liquid adding device includes a washing liquid, Three kinds of reagents and substrates for capturing antibodies or antigens.
  • the number of the tanks on the mounting frame is set to four, wherein the reagents in the liquid reagent cups in the two tanks are washed, and the reagents in the liquid reagent cup in the other tank are used to capture antibodies or antigens.
  • the reagent, the reagent in the liquid reagent cup in the last tank, uses a substrate.
  • the capillary body is made of glass, quartz or polystyrene, and the capillary body has a tube length of 20 to 100 mm and an inner diameter of 0.2 to 1 mm.
  • the sealing film is composed of an aluminum plastic film or a polytetrafluoroethylene film.
  • the bottom end of the tank body is further provided with a convex member, the piercing device is fixed at the top end of the convex member; the height of the groove body is L, the convex member The height is a, the height of the puncture device is b, and the height of the liquid reagent cup is c, then L ⁇ a+b+c.
  • the shape of the liquid reagent cup matches the shape of the tank.
  • a capillary chemiluminescence detection method comprising:
  • the antibody or antigen in the sample solution reacts with the antibody or antigen coated in the capillary body, and after the reaction is completed, the top end of the capillary body is blown to discharge the liquid in the capillary body;
  • step (3) repeating step (3) to perform cleaning of the capillary body
  • the residue inside the capillary body can be effectively cleaned and the detection accuracy can be improved.
  • the present invention has the following advantages and beneficial effects:
  • the invention can perform chemiluminescence reaction on a separate capillary tube, reduce the amount of reagents in the immune reaction while avoiding contamination of the reagent source, and realize independent packaging of the single reagent, which can effectively simplify the reagent in the immune experiment. Measure the work and maximize the reduction of the existing human error, reduce the workload and improve the stability of the test;
  • the invention simultaneously utilizes the small volume of the capillary itself and the principle of capillary action to realize the control of the reaction volume and reduces the requirement for the accuracy of the sample loading.
  • the invention not only reduces the cost of the instrument, but also improves the operation. reliability.
  • Figure 1 is a schematic view of the structure of the present invention.
  • Figure 2 is a schematic view of the internal structure of the present invention.
  • Figure 3 is a schematic cross-sectional view showing the liquid reagent cup of the present invention.
  • a capillary chemiluminescence detecting device comprises a liquid adding device for adding a reagent to the capillary body 1.
  • the capillary body 1 is made of a glass material, and the capillary body 1 has a tube length of 40 mm and an inner diameter of 0.6. Millimeter.
  • the liquid adding device comprises a funnel cup 2, a mounting frame 3, a tank body, a liquid reagent cup 4 and a puncture device 6, and the specific structure is as follows:
  • the bottom end of the funnel cup 2 is connected with the top end of the capillary body 1, and the mounting frame 3 is fixed at the top end of the funnel cup 2, and the mounting frame 3 is provided
  • the tank body is disposed, the bottom end of the tank body is connected with the top end of the funnel cup 2, the puncture device 6 is fixed at the bottom end of the tank body, and the liquid reagent cup 4 is placed in the tank body.
  • the invention also optimizes the structure of the liquid reagent cup 4, and promotes the effective flow of the reagent in the liquid reagent cup 4 into the capillary body 1 during the pressing process of the liquid reagent cup 4.
  • the specific arrangement is as follows: the liquid reagent cup 4 is a cup 41 having an opening, a reagent disposed in the cup 41, and a sealing film 42 disposed at the opening of the cup 41; the opening of the liquid reagent cup 4 is downward, during the depression of the cup 41
  • the puncture device 6 can pierce the sealing film 42 and release the liquid sealed in the liquid reagent cup 4 into the tank body, and then flow into the capillary body 1 through the funnel cup 2.
  • the number of the tanks on the mounting frame 3 is preferably set to four, wherein the reagents in the liquid reagent cup 4 in the two tanks are washed, and the reagents in the liquid reagent cup 4 in the other tank are trapped antibodies. Or the reagent of the antigen, the reagent in the liquid reagent cup 4 in the last tank uses the substrate, as shown in Figures 1-3.
  • the specific detection method of the above capillary chemiluminescence detecting device is as follows:
  • the mechanical device will be used to grasp the liquid adding device with the capillary body 1 to the sample suction position, and at the same time, the vacuum suction nozzle similar to the pipette is lowered to be tightly combined with the inner wall of the funnel cup 2, when the lower end of the capillary body 1 is immersed
  • the internal vacuum device of the supporting instrument starts to extract the negative pressure, and the sample solution is sucked into the capillary body; then the vacuum suction head is separated from the inner wall of the funnel cup 2, and then the mechanical device is used to grasp the liquid adding device to the reaction zone. Carry out the reaction;
  • the solution is sampled by a mechanical device.
  • the reaction zone is usually thermostated at 37 degrees. Under this condition, the antibody or antigen in the sample solution is first reacted with the antibody or antigen coated in the capillary body, and the capillary body is manually used after the reaction is completed. The liquid is completely blown out, or the automatic blowing mechanism of the supporting instrument is moved down to the top of the funnel cup, and the top end of the capillary body is blown by the automatic blowing mechanism to discharge the liquid in the capillary body;
  • the capillary liquid release mechanism of the manual or supporting instrument is used to press the liquid reagent cup containing the washing liquid to wash the capillary body, and after the cleaning is completed, the automatic blowing mechanism of the manual or supporting instrument is used to blow the capillary body.
  • a capillary liquid release mechanism using a manual or an accessory device is used to press a liquid reagent cup containing a reagent for capturing an antibody or an antigen, and a reagent for capturing an antibody or an antigen flows along a slope between the liquid reagent cup and the funnel cup to the funnel cup.
  • the reagent in the funnel cup flows into the inside of the capillary body under the action of its own gravity and the capillary body of the capillary body, so that the reagent for capturing the antibody or the antigen reacts with the substance in the capillary body for a second reaction, and the reaction is completed by manual or matching.
  • the automatic blowing mechanism of the instrument blows out the liquid in the capillary body;
  • step (3) repeating step (3) to perform cleaning of the capillary body
  • the capillary liquid release mechanism of the manual or supporting instrument is used to press down the liquid reagent cup containing the substrate, and the substrate is released and flows into the capillary body to react the substrate with the substance in the capillary body, and then reacted.
  • the capillary can be optically detected.
  • the material of the capillary body (1) and the sealing film (42) is optimized in this embodiment.
  • the capillary body (1) is made of a polystyrene material.
  • the sealing film (42) is made of an aluminum plastic film.
  • the bottom end of the trough body is further provided with a convex member 5, the puncture device 6 is fixed at the top end of the convex member 5; the height of the trough body is L, the height of the convex member 5 is a, the height of the puncture device 6 is b, the liquid reagent cup The height of 4 is c, then L ⁇ a + b + c.
  • the shape of the liquid reagent cup (4) matches the shape of the tank.
  • the capillary liquid release mechanism of the manual or supporting instrument is used to press the liquid reagent cup containing the washing liquid, and the sealing film of the liquid reagent cup is squeezed, and the liquid reagent cup itself is pressed for about one third of the height, and then the pressing is stopped.
  • the washing liquid is flushed into the capillary body and reaches a stable state
  • the liquid in the capillary body is completely blown out by an automatic blowing mechanism of a manual or an auxiliary instrument, and then the liquid reagent cup itself is pressed down by about one-third of the height of the liquid reagent cup.
  • the liquid in the capillary body After flushing into the capillary body and reaching the steady state again, the liquid in the capillary body is completely blown out by the automatic blowing mechanism of the manual or the matching instrument, and finally the liquid reagent cup is completely pressed to make the liquid out, when the washing liquid is washed in.
  • the liquid in the capillary body can be completely blown out by using an automatic blowing mechanism of a manual or an accessory instrument.

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Abstract

一种毛细管化学发光检测装置及其检测方法，解决了现有技术存在成本高、工作稳定性不足、液体源污染的问题。装置包括用于向毛细管本体（1）中添加试剂的加液装置；加液装置包括底端与毛细管本体（1）的顶端连通的漏斗杯（2），具有一个以上槽体的安装架（3），设置在每个槽体内用于盛装试剂的液体试剂杯（4）；液体试剂杯（4）由具有开口的杯体（41），设置在杯体（41）内的试剂，以及设置在杯体（41）的开口处的密封薄膜（42）组成；槽体的底端与漏斗杯（2）的顶端连通，槽体的内部底端还设置有用于液体试剂杯（4）下压时刺穿密封薄膜（42）使液体试剂杯（4）内试剂流入漏斗杯（2）中的穿刺装置（6）。具有既降低了仪器的成本，又提高了运行的可靠性等优点。

Description

一种毛细管化学发光检测装置及其检测方法 技术领域
本发明属于化学发光检测领域，具体涉及一种毛细管化学发光检测装置及其检测方法。
背景技术
一次性点样毛细管具有自动吸取液体的毛细现象，能够实现自动加液，但仅限于第一次使用时有效，当使用过一次后同一根毛细管再次利用毛细现象加液则会造成后面使用的液体源的污染。这样利用毛细管作为一次性反应容器时，就无法进行像免疫实验之类的多步骤反应，免疫实验过程中要使用2种抗体或抗原对待测抗原或抗体进行结合，同时还会使用到洗液和底物等通用试剂辅助实验进行。如果作为固定反应容器，则需要增加清洗装置及清洗过程。
近几年兴起的微流控芯片，有部分产品也是利用了毛细管来进行免疫反应，作为反应的容器，如CN201510696773.2，一般利用毛细管作为微流控反应容器或通道的产品都会将全部液体试剂整合到芯片内，做到单人份的独立包装，但这类产品都仅是利用了毛细管体积小的特点，将试剂量降到最小，且毛细管仅作为反应通道或容器使用。而为了实现精确的定量反应，各试剂整合到芯片上，所采用的动力系统和液体加样系统都很复杂，必须能够实现相当高的精度才行，因此此类微流控芯片的仪器与耗材价格都会居高不下，且整个系统工作稳定性也会大幅下降。
发明内容
本发明所要解决的技术问题是：现有技术中利用毛细管作为一次性反应容器时无法进行像免疫实验之类的多步骤反应，而能够利用毛细管作为免疫反应的容器则存在成本高、工作稳定性不足、液体源污染的问题，目的在于提供了一种毛细管化学发光检测装置及其检测方法，其不仅仅能有效利用毛细管作为一次性反应容器，降低试剂用量，并且能有效解决试剂源的污染问题，还能利用了毛细管本身的小体积来实现对反应体积的控制，降低对加样精度的需求，本发明既降低了仪器的成本，又提高了运行的可靠性。
本发明通过下述技术方案实现：
一种毛细管化学发光检测装置，包括用于向毛细管本体中添加试剂的加液装置；所述加液装置包括底端与毛细管本体的顶端连通的漏斗杯，具有一个以上槽体的安装架，设置在每个槽体内用于盛装试剂的液体试剂杯；所述液体试剂杯由具有开口的杯体，设置在杯体内的试剂，以及设置在杯体的开口处的密封薄膜组成；所述槽体的底端与漏斗杯的顶端连通，所 述槽体的内部底端还设置有用于液体试剂杯下压时刺穿密封薄膜使液体试剂杯内试剂流入漏斗杯中的穿刺装置。
本发明的液体试剂杯开口向下倒置于加液装置内，在液体试剂杯下方对应的加液装置上设计有便于穿破密封薄膜的穿刺装置，当需要加入某种液体试剂时，人工或配套仪器上的运动机构下压，使穿刺装置刺破液体试剂杯的密封薄膜，继续挤压，液体流出。由于穿刺装置位置高于毛细管本体上端的开口，液体在重力作用下流到毛细管本体的上端开口处，再在毛细管本体的毛细作用与液体本身的重力共同作用下，流入毛细管本体内部。液体所受重力大于毛细作用力时，液体会从毛细管下口流出，液体减少，直到其所受重力与毛细作用力相等时，停止流出，在毛细管内保持稳定状态。当液体试剂反应或清洗结束，采用人工或配套仪器上的毛细管液体释放机构下压，吹出压力较大的气体，将毛细管中的液体从下口吹出，落入底部废液收集器中即可，吹气过程持续2s-10s，可有效确保将液体全部吹出。
同时，毛细管具有毛细现象，毛细现象是在一些线度小到足以与液体弯月面的曲率半径相比较的毛细管中发生的现象，毛细管中整个液体表面都将变得弯曲，液固分子间的相互作用可扩展到整个液体，液体表面类似张紧的橡皮膜，如果液面是弯曲的，它就有变平的趋势，因此凹液面对下面的液体施以拉力，凸液面对下面的液体施以压力。如果浸润液体在毛细管中的液面是凹形的，它对下面的液体施加拉力，使液体沿着管壁上升，当向上的拉力跟管内液柱所受的重力相等时，管内的液体停止上升，达到平衡。
因而，本发明将一次性反应容器毛细管本体与带有多种液体试剂杯的加液装置整合到一起，并使液体试剂按单次反应需要量独立封装到液体试剂杯内后，只需按后续实验流程依次挤破液体试剂杯，将液体释放流入到毛细管本体内进行反应，即可实现在一根独立的毛细管上进行化学发光反应的目的。上述方式既可以解决一次性毛细管加入多种试剂时对试剂源的污染问题，又可以省略免疫反应试剂在免疫反应时的量取操作，减轻了免疫反应试剂量取的劳动强度，并且避免了量取过程中存在的人为误差，提高了检测的稳定性。
同时，本发明还利用了毛细管本身的小体积来实现对反应体积的控制，不仅仅能达到现有技术中采用毛细管结构能够减少检测时试剂量的作用，而且，通过本发明结构的优化和与毛细现象的原理相结合后，还能有效降低对加样精度的需求，因而，本发明与现有技术相比既降低了仪器的成本，又提高了运行的可靠性。
进一步，所述毛细管本体内包被有抗体或抗原，所述安装架上槽体的数量设置为三个以上，每个液体试剂杯中采用一种试剂，加液装置中的试剂包括有洗液、捕获抗体或抗原的试剂、底物三种。
优选地，所述安装架上槽体的数量设置为四个，其中两个槽体内的液体试剂杯中的试剂采用洗液，另一个槽体内的液体试剂杯中的试剂采用捕获抗体或抗原的试剂，最后一个槽体内的液体试剂杯中的试剂采用底物。
更进一步地，所述毛细管本体由玻璃、石英或聚苯乙烯材质构成，毛细管本体的管长为20～100毫米，内径为0.2～1毫米。所述密封薄膜采用铝塑薄膜或聚四氟乙烯薄膜构成。
为了能更好地达到将液体试剂杯中液体流入到毛细管本体中，所述槽体底端还设置有凸件，该穿刺装置固定在凸件顶端；设槽体的高度为L，凸件的高度为a，穿刺装置的高度为b，液体试剂杯的高度为c，则L≥a+b+c。通过上述结构的设置，还能有效实现液体试剂杯内试剂分批次流入到毛细管本体中。
并且，通过上述设置，不仅仅能有效便于液体试剂杯的下压操作，使液体试剂杯中的试剂更好地流入毛细管本体内，而且还能有效避免液体试剂杯的上表面高于槽体上表面，进而减少外力意外造成的液体试剂杯上密封薄膜被穿刺装置破坏的情况发生。
进一步，所述液体试剂杯的形状与槽体的形状相匹配。通过该结构的优化设置，能有效促液体试剂杯下压过程中，该液体试剂杯内试剂能完全压出，进而更好地保证试剂能流入到毛细管本体中，提高检测稳定性。
一种毛细管化学发光检测方法，包括：
(1)将样本溶液加入到毛细管本体中；
(2)样本溶液中的抗体或抗原与包被在毛细管本体中的抗体或抗原反应，反应完成后对毛细管本体的顶端吹气，使毛细管本体中的液体排出；
(3)下压盛装有洗液的液体试剂杯，使洗液冲洗毛细管本体，清洗完成后，用毛细管液体释放机构吹出毛细管本体中的液体；
(4)下压盛装有捕获抗体或抗原的试剂的液体试剂杯，使捕获抗体或抗原的试剂流入毛细管本体中进行反应，反应完成后吹出毛细管本体中的液体；
(5)重复步骤(3)进行毛细管本体的清洗；
(6)下压盛装有底物的液体试剂杯，将底物释放并流入到毛细管本体内，底物与毛细管本体内物质反应后进行检测即可。
进一步，所述步骤(3)中洗液冲洗毛细管本体的具体步骤如下：
下压将盛装有洗液的液体试剂杯的密封薄膜挤破，压下全部行程的三分之一，当洗液冲入毛细管本体并达到稳定状态后，将毛细管本体内的液体全部吹出，然后再下压行程的三分之一，当洗液冲入毛细管本体并达到稳定状态后，再将毛细管本体内的液体全部吹出，然后 再全部压下液体试剂杯使液体全部压出，当洗液冲入毛细管本体并达到稳定状态后，最后再将毛细管本体内的液体全部吹出。
通过上述步骤(3)的优化能有效确保毛细管本体内部的残留物冲洗干净，提高检测准确度。
本发明与现有技术相比，具有如下的优点和有益效果：
1、本发明能在一根独立的毛细管上进行化学发光反应，在减少免疫反应中试剂用量的同时避免对试剂源的污染，并且实现了单份试剂的独立包装，能有效简化免疫实验中试剂的量取工作，并最大化的减少量取存在的人为误差，减轻工作量的同时提高了检测的稳定性；
2、本发明同时利用了毛细管本身的小体积以及毛细作用的原理来实现了对反应体积的控制，并降低了对加样精度的需求，本发明既降低了仪器的成本，又提高了运行的可靠性。
附图说明
此处所说明的附图用来提供对本发明实施例的进一步理解，构成本申请的一部分，并不构成对本发明实施例的限定。在附图中：
图1为本发明的结构示意图。
图2为本发明的内部结构示意图。
图3为本发明中液体试剂杯的剖面结构示意图。
附图中标记及对应的零部件名称：
1-毛细管本体，2-漏斗杯，3-安装架，4-液体试剂杯，5-凸件，6-穿刺装置；
41-杯体，42-密封薄膜。
具体实施方式
为使本发明的目的、技术方案和优点更加清楚明白，下面结合实施例和附图，对本发明作进一步的详细说明，本发明的示意性实施方式及其说明仅用于解释本发明，并不作为对本发明的限定。
实施例1
一种毛细管化学发光检测装置，包括用于向毛细管本体1中添加试剂的加液装置，本实施例中该毛细管本体1由玻璃材质构成，该毛细管本体1的管长为40毫米，内径为0.6毫米。
该加液装置包括漏斗杯2、安装架3、槽体、液体试剂杯4和穿刺装置6，具体结构如下：
漏斗杯2底端与毛细管本体1顶端连通，漏斗杯2顶端固定有安装架3，安装架3上设 置有槽体，槽体底端与，漏斗杯2顶端连通，穿刺装置6固定在槽体底端，液体试剂杯4放置在槽体内。
同时，本发明还优化了液体试剂杯4的结构，促使在液体试剂杯4下压过程中能有效将液体试剂杯4中的试剂流入毛细管本体1中，具体设置方式为：液体试剂杯4由具有开口的杯体41，设置在杯体41内的试剂，以及设置在杯体41的开口处的密封薄膜42组成；所述液体试剂杯4的开口向下，在杯体41下压过程中，穿刺装置6即可刺破密封薄膜42，进而将密封在液体试剂杯4内的液体释放到槽体内，进而通过漏斗杯2流入到毛细管本体1中。
本实施例中安装架3上槽体的数量优选设置为四个，其中两个槽体内的液体试剂杯4中的试剂采用洗液，另一个槽体内的液体试剂杯4中的试剂采用捕获抗体或抗原的试剂，最后一个槽体内的液体试剂杯4中的试剂采用底物，如图1-图3所示。
上述一种毛细管化学发光检测装置的具体检测方法如下：
(1)人工用移液器将适量样本从图1所示的漏斗杯2位置处加入，然后放入反应区进行反应；
或将采用机械装置将带有毛细管本体1的加液装置抓取到样本吸入位，同时采用类似于移液器的真空吸头下降，与漏斗杯2的内壁紧密结合，当毛细管本体1下端浸入到样本溶液后，配套仪器的内部真空装置启动抽取负压，将样本溶液吸入到毛细管本体中；然后真空吸头与漏斗杯2的内壁脱离，再由机械装置将加液装置抓取到反应区进行反应；
本实施例中采用机械装置吸取样本溶液。
(2)该反应区通常恒温37度，在该条件下，样本溶液中的抗体或抗原与包被在毛细管本体中的抗体或抗原进行第一次反应，反应完成后采用人工将毛细管本体内的液体全部吹出，或配套仪器的自动吹气机构下移到漏斗杯顶端，采用自动吹气机构对毛细管本体的顶端吹气，使毛细管本体中的液体排出；
(3)采用人工或配套仪器的毛细管液体释放机构下压盛装有洗液的液体试剂杯，使洗液冲洗毛细管本体，清洗完成后，采用人工或配套仪器的自动吹气机构吹出毛细管本体中的液体；
(4)采用人工或配套仪器的毛细管液体释放机构下压盛装有捕获抗体或抗原的试剂的液体试剂杯，捕获抗体或抗原的试剂沿着液体试剂杯与漏斗杯之间的斜面流入到漏斗杯内，漏斗杯内的试剂在自身重力及毛细管本体的毛细作用下，流入到毛细管本体内部，使捕获抗体或抗原的试剂与毛细管本体中的物质进行第二次反应，反应完成后采用人工或配套仪器的自动吹气机构吹出毛细管本体中的液体；
(5)重复步骤(3)进行毛细管本体的清洗；
(6)采用人工或配套仪器的毛细管液体释放机构下压盛装有底物的液体试剂杯，将底物释放并流入到毛细管本体内，使底物与毛细管本体内物质反应后，将反应后的毛细管进行光学检测即可。
实施例2
本实施例与实施例1的区别在于，本实施例对毛细管本体(1)和密封薄膜(42)的材质进行了优化，本实施例中毛细管本体(1)由聚苯乙烯材质构成。该密封薄膜(42)采用铝塑薄膜构成。
实施例3
本实施例与实施例1的区别在于，本实施例中优化了槽体位置处的结构，如图2所示，具体设置如下：
该槽体底端还设置有凸件5，该穿刺装置6固定在凸件5顶端；设槽体的高度为L，凸件5的高度为a，穿刺装置6的高度为b，液体试剂杯4的高度为c，则L≥a+b+c。液体试剂杯(4)的形状与槽体的形状相匹配。
本实施例中该槽体的高度值L等于凸件5的高度a、穿刺装置6的高度b和液体试剂杯4的高度c之和；即，L＝a+b+c。
实施例4
本实施例与实施例1的区别在于，本实施例中步骤(3)和(5)的清洗过程不同，具体设置如下：
采用人工或配套仪器的毛细管液体释放机构下压盛装有洗液的液体试剂杯，将液体试剂杯的密封薄膜挤破，压下液体试剂杯本身高度的三分之一左右后停止下压，当洗液冲入毛细管本体并达到稳定状态后，采用人工或配套仪器的自动吹气机构将毛细管本体内的液体全部吹出，然后再下压液体试剂杯本身高度的三分之一左右，当洗液冲入毛细管本体并再次达到稳定状态后，又采用人工或配套仪器的自动吹气机构将毛细管本体内的液体全部吹出，最后再全部压下液体试剂杯使液体全部压出，当洗液冲入毛细管本体并最后一次在毛细管本体内达到稳定状态后，采用人工或配套仪器的自动吹气机构将毛细管本体内的液体全部吹出即可。通过本实施例中该方式的设置，能有效增加清洗效果。
以上所述的具体实施方式，对本发明的目的、技术方案和有益效果进行了进一步详细说明，所应理解的是，以上所述仅为本发明的具体实施方式而已，并不用于限定本发明的保护范围，凡在本发明的精神和原则之内，所做的任何修改、等同替换、改进等，均应包含在本 发明的保护范围之内。

Claims (10)

  1. 一种毛细管化学发光检测装置，其特征在于，包括用于向毛细管本体(1)中添加试剂的加液装置；所述加液装置包括底端与毛细管本体(1)的顶端连通的漏斗杯(2)，具有一个以上槽体的安装架(3)，设置在每个槽体内用于盛装试剂的液体试剂杯(4)；所述液体试剂杯(4)由具有开口的杯体(41)，设置在杯体(41)内的试剂，以及设置在杯体(41)的开口处的密封薄膜(42)组成；所述槽体的底端与漏斗杯(2)的顶端连通，所述槽体的内部底端还设置有用于液体试剂杯(4)下压时刺穿密封薄膜(42)使液体试剂杯(4)内试剂流入漏斗杯(2)中的穿刺装置(6)。
  2. 根据权利要求1所述的一种毛细管化学发光检测装置，其特征在于，所述毛细管本体(1)包被有抗体或抗原，所述安装架(3)上槽体的数量设置为三个以上，每个液体试剂杯(4)中采用一种试剂，加液装置中的试剂包括有洗液、捕获抗体或抗原的试剂、底物三种。
  3. 根据权利要求2所述的一种毛细管化学发光检测装置，其特征在于，所述安装架(3)上槽体的数量设置为四个，其中两个槽体内的液体试剂杯(4)中的试剂采用洗液，另一个槽体内的液体试剂杯(4)中的试剂采用捕获抗体或抗原的试剂，最后一个槽体内的液体试剂杯(4)中的试剂采用底物。
  4. 根据权利要求1所述的一种毛细管化学发光检测装置，其特征在于，所述毛细管本体(1)由玻璃、石英或聚苯乙烯材质构成。
  5. 根据权利要求1所述的一种毛细管化学发光检测装置，其特征在于，所述毛细管本体(1)的管长为20～100毫米，内径为0.2～1毫米。
  6. 根据权利要求1所述的一种毛细管化学发光检测装置，其特征在于，所述密封薄膜(42)采用铝塑薄膜或聚四氟乙烯薄膜构成。
  7. 根据权利要求1-6任一项所述的一种毛细管化学发光检测装置，其特征在于，所述槽体底端还设置有凸件(5)，该穿刺装置(6)固定在凸件(5)顶端；设槽体的高度为L，凸件(5)的高度为a，穿刺装置(6)的高度为b，液体试剂杯(4)的高度为c，则L≥a+b+c。
  8. 根据权利要求7所述的一种毛细管化学发光检测装置，其特征在于，所述液体试剂杯(4)的形状与槽体的形状相匹配。
  9. 一种毛细管化学发光检测方法，其特征在于，包括：
    (1)将样本溶液加入到毛细管本体中；
    (2)样本溶液中的抗体或抗原与包被在毛细管本体中的抗体或抗原反应，反应完成后对毛细管本体的顶端吹气，使毛细管本体中的液体排出；
    (3)下压盛装有洗液的液体试剂杯，使洗液冲洗毛细管本体，清洗完成后，用毛细管液体释放机构吹出毛细管本体中的液体；
    (4)下压盛装有捕获抗体或抗原的试剂的液体试剂杯，使捕获抗体或抗原的试剂流入毛细管本体中进行反应，反应完成后吹出毛细管本体中的液体；
    (5)重复步骤(3)进行毛细管本体的清洗；
    (6)下压盛装有底物的液体试剂杯，将底物释放并流入到毛细管本体内，底物与毛细管本体内物质反应后进行检测即可。
  10. 根据权利要求9所述的一种毛细管化学发光检测方法，其特征在于，所述步骤(3)中洗液冲洗毛细管本体的具体步骤如下：
    下压将盛装有洗液的液体试剂杯的密封薄膜挤破，压下全部行程的三分之一，当洗液冲入毛细管本体并达到稳定状态后，将毛细管本体内的液体全部吹出，然后再下压行程的三分之一，当洗液冲入毛细管本体并达到稳定状态后，再将毛细管本体内的液体全部吹出，然后再全部压下液体试剂杯使液体全部压出，当洗液冲入毛细管本体并达到稳定状态后，最后再将毛细管本体内的液体全部吹出。
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