WO2024021664A1 - 一种可变色指示的免疫层析试纸条及其使用方法 - Google Patents

一种可变色指示的免疫层析试纸条及其使用方法 Download PDF

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Publication number
WO2024021664A1
WO2024021664A1 PCT/CN2023/085581 CN2023085581W WO2024021664A1 WO 2024021664 A1 WO2024021664 A1 WO 2024021664A1 CN 2023085581 W CN2023085581 W CN 2023085581W WO 2024021664 A1 WO2024021664 A1 WO 2024021664A1
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pad
sample
absorbent pad
color
test strip
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PCT/CN2023/085581
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English (en)
French (fr)
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陈亚茹
赵弟萍
倪建良
杨姗姗
张国锋
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厦门宝太生物科技股份有限公司
杭州宝太生物科技有限公司
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Publication of WO2024021664A1 publication Critical patent/WO2024021664A1/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
    • 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
    • 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
    • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
    • G01N21/78—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a change of colour

Definitions

  • the invention relates to a color-changing indicator immunochromatography test strip and a method of using the same, and belongs to the technical field of immunochromatography.
  • Immunochromatography is a pair of specific antigens or antibodies, one of which is fixed on a nitrocellulose membrane, the other is labeled with a tracer and made into a binding pad, and finally assembled into a test strip; in the test
  • the sample and diluent are added to the adding end of the paper strip, which is the position of the sample pad.
  • the sample will chromatograph along the nitrocellulose membrane due to the chromatographic force. If there is a substance to be tested in the sample, the substance to be tested will be When combined with the antigen-antibody with tracer in the binding pad, it will be intercepted by the antigen-antibody immobilized on the nitrocellulose membrane and display a certain color, thereby achieving specific immunodiagnosis.
  • Immunochromatography has been widely used in disease diagnosis, eugenics, food testing, environmental monitoring and other fields because it is simple, convenient, fast, cheap and does not require the operation of large instruments and professionals.
  • the immunochromatography test strips commonly used in immunochromatography mainly include five parts: sample addition area (sample pad), labeling area (binding pad), display area (nitrocellulose membrane), and water-absorbent area (absorbent pad) and backing board.
  • the water-absorbent area is composed of conventional untreated water-absorbent materials such as AN3, AN5 and other filter papers, which provide a certain chromatographic force for chromatography; the internal quality control system of the product is the C system.
  • C line quality control line
  • the invention provides a color-changing indicator immunochromatography test strip and a method of using the same, which can effectively solve the above problems.
  • the present invention is implemented as follows:
  • An immunochromatography test strip with color changeable indication including a bottom plate and a sample pad, a binding pad, a reaction membrane and a water-absorbent pad connected sequentially along the chromatographic direction on the bottom plate.
  • a detection line is set on the reaction membrane and no quality control is provided. Line; the water-absorbent pad changes color when the detection sample liquid arrives.
  • the water-absorbent pad is a water-absorbent pad treated with an acid-base indicator.
  • the acid-base indicator treatment involves soaking the absorbent pad in the acid-base indicator treatment solution or applying the acid-base indicator treatment solution on the absorbent pad, and then drying it for use after a period of time.
  • it also includes a shell covering the immunochromatography test strip, and the shell is provided with a sample addition area for dripping detection sample liquid, a detection area for observing or reading signals, and An indication area is used to determine whether the running board is effective.
  • the sample adding area is located above the sample pad, the detection area is located above the detection line, and the indication area is located above the absorbent pad.
  • a method of using the above-mentioned color-changing indicator immunochromatography test strip includes the following steps:
  • an alkaline buffer or an acidic buffer is added to the detection sample solution before dropwise addition.
  • the present invention applies the color-changing absorbent pad to the immunochromatography test paper without using the independent protein system of the traditional C system, thereby avoiding cross-interference during the chromatography process and achieving high specificity.
  • the present invention applies the color-changing absorbent pad to the immunochromatography test paper.
  • the color-changing absorbent pad is cheaper, and the reading results are quick and intuitive. It does not require special instruments and equipment, nor does it require professional operation. , which can save a lot of material costs and labor operation costs.
  • Figure 1 is a schematic structural diagram of a color-changing indicator immunochromatography test strip provided by an embodiment of the present invention.
  • Figure 2 is a schematic diagram of the shell structure of a color-changeable indicator immunochromatography test strip provided by an embodiment of the present invention.
  • Sample pad 1 binding pad 2, reaction membrane 3, detection line 31, absorbent pad 4, bottom plate 5, sample addition area 6, detection area 7, indicator area 8.
  • first and second are used for descriptive purposes only and cannot be understood as indicating or implying relative importance or implicitly indicating the number of indicated technical features. Therefore, features defined as “first” and “second” may explicitly or implicitly include one or more of these features.
  • “plurality” means two or more than two, unless otherwise explicitly and specifically limited.
  • an embodiment of the present invention provides a color-changing indicator immunochromatography test strip, including a bottom plate 5 and a sample pad 1, a binding pad 2, a reaction membrane 3 and a Absorbent pad 4.
  • a detection line 31 is provided on the reaction membrane 3, and no quality control line is provided.
  • the water-absorbent pad 4 changes color when the detection sample liquid arrives. If the water-absorbent pad 4 changes color after the detection is completed, it means that the sample liquid used for detection has reached the water-absorbent pad 4, and must have passed the detection line 31 set on the reaction membrane 3, and it can be judged that the running board is effective; otherwise, The running board is invalid and the running board needs to be tested again.
  • the water-absorbent pad 4 is a water-absorbent pad treated with an acid-base indicator.
  • the test sample liquid is generally acidic or alkaline. When the test sample liquid reaches the absorbent pad, the acid-base indicator on the absorbent pad will change color, indicating that the running board is effective.
  • the acid-base indicator treatment involves soaking the absorbent pad in the acid-base indicator treatment solution or applying the acid-base indicator treatment solution on the absorbent pad, and then drying it for use after a period of time.
  • the formula of the acid-base indicator treatment solution is 53wt% ethanol, 0.0047wt% methyl red, 0.10wt% bromothymol blue, 0.45wt% 20wt% Tween20, 18wt% 5wt%PVA, 19wt% 5wt%PVP solution.
  • the color-changing indicator immunochromatography test strip further includes a shell covering the immunochromatography test strip, and the shell is provided with a sample addition area for dripping the detection sample solution. 6.
  • the sample adding area 6 is located above the sample pad 1, and the detection area 7 is located above the detection line 31.
  • the indication area 8 is located above the water-absorbent pad 4 .
  • the length ratio of the sample pad 1, the binding pad 2, the reaction membrane 3, and the water-absorbent pad 4 is 2:1:4:1; this length ratio is convenient for adding samples to the sample pad 1 and reacting at the same time.
  • the membrane 3 is long enough to allow the sample liquid to stay on the reaction membrane 3 for a sufficient time to complete the detection reaction.
  • the sample pad 1 and the binding pad 2 overlap at the connection point, and the sample pad 1 overlaps above the binding pad 2.
  • the length of the overlapping portion is 1/4-1/2 of the length of the binding pad 2, which is convenient for sample liquid Flow smoothly from the sample pad 1 to the binding pad 2.
  • the bonding pad 2 and the reaction membrane 3 overlap at the connection point, and the bonding pad 2 overlaps above the reaction membrane 3.
  • the length of the overlapping portion is 1/8-1/4 of the length of the reaction membrane 3; it is convenient for sample liquid Flows smoothly from the bonding pad 2 to the reaction membrane 3 .
  • the reaction film 3 and the water-absorbent pad 4 overlap at the connection point, and the water-absorbent pad 4 overlaps above the reaction film 3.
  • the length of the overlapping portion is 1/5-1/3 of the length of the water-absorbent pad 4, which facilitates the smooth flow of the sample liquid. It is adsorbed from the reaction film 3 to the water-absorbing pad 4 .
  • the detection line 31 is located at 1/2 of the reaction film 3 .
  • Such a size setting facilitates the sample liquid chromatography to flow along the direction of the sample pad 1, binding pad 2, reaction membrane 3, and water-absorbent pad 4. At the same time, the sample liquid stays on the reaction membrane 3 for enough time to complete the detection reaction, improving Detection sensitivity and efficiency.
  • the sample pad 1 is a glass fiber membrane or a polyester fiber membrane.
  • the bonding pad 2 is a glass fiber membrane or a polyester fiber membrane.
  • reaction membrane 3 is a nitrocellulose membrane.
  • the water-absorbent pad 4 is filter paper (such as AN3, AN5)
  • the base plate 5 is a PVC base plate or a PS base plate.
  • the test sample liquid is one or more of serum, whole blood, urine, tissue fluid, throat swabs, nasal swabs, oropharyngeal swabs, anal swabs or excreta.
  • An embodiment of the present invention also provides a method for using the above-mentioned color-changing indicator immunochromatography test strip, which includes the following steps:
  • the detection sample liquid dropwise on the sample pad 1, the amount of the dropwise addition is 1-3 drops, and let it sit for a period of time, usually 10-20 minutes.
  • the detection sample liquid is along the sample pad 1, the binding pad 2, the reaction membrane 3 and Chromatography on the absorbent pad 4, the detection is completed when the chromatography reaches the absorbent pad;
  • the detection result is read by observing the detection line 31.
  • an alkaline buffer or an acidic buffer is added to the detection sample solution before dropwise addition.
  • the pH of the alkaline buffer or acidic buffer can be adjusted according to the type of acid-base indicator treatment solution. It is further preferred to add an alkaline buffer solution.
  • the detection specificity of the color-changing indicator immunochromatographic test strip of Example 1 was tested.
  • the test method is to use many negative samples to conduct detection experiments to see whether there will be false positive samples, so as to calculate the specificity.
  • the immunochromatography test strips of the traditional protein C-line system were used as the control group, and the color-changing indicator immunochromatography test paper strips of Example 1 were used as the experimental group. In both groups, 200 cases of serum, 200 cases of plasma, and 200 cases of whole blood were collected. The statistical experimental results are shown in Table 1.
  • the detection specificity of the color-changing indicator immunochromatographic test strip of Example 1 was tested.
  • the test method is to use many negative samples to conduct detection experiments to see whether there will be false positive samples, so as to calculate the specificity.
  • the immunochromatography test strips of the traditional protein C-line system were used as the control group, and the color-changing indicator immunochromatography test paper strips of Example 1 were used as the experimental group. 500 negative serum samples were taken from both groups. The statistical experimental results are shown in Table 2.
  • the immunochromatography test strip of the traditional protein C-line system was used as the control group, and the color-changing indicator immunochromatography test paper strip of Example 1 was used as the experimental group. Comparison The backgrounds of both ran completely clean. Criteria for judging whether the background is completely clean: there is no obvious red background in the window. The statistical results are shown in Table 3.
  • Group group background clean time () () (S) sample book 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15 control group 90s 89s 93s 88s 90s 91s 95s 87s 80s 85s 89s 91s 92s 91s 89s test group 80s 79s 78s 77s 78s 79s 76s 79s 75s 76s 75s 73s 75s 74s 76s
  • Table 3 the background clearing time of the color-changing indicator immunochromatography test strip of Example 1 is shorter, indicating that its detection efficiency is higher.
  • the analysis reason is that the color-changing indicator immunochromatography test paper strip of Example 1 of the present invention does not have a quality control line, which reduces one reaction procedure, and the detection sample solution stays on the reaction membrane for a shorter time, thereby shortening the The entire background cleaning time improves detection efficiency.

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Abstract

本发明提供一种可变色指示的免疫层析试纸条及其使用方法，包括底板和底板上沿层析方向依次连接的样品垫、结合垫、反应膜及吸水垫，其特征在于，所述反应膜上设置检测线，不设置质控线；所述吸水垫在检测样品液到达时变色。所述可变色指示的免疫层析试纸条的检测特异性高，成本低。

Description

一种可变色指示的免疫层析试纸条及其使用方法 技术领域
本发明涉及一种可变色指示的免疫层析试纸条及其使用方法，属于免疫层析技术领域。
 背景技术
免疫层析法(immunochromatography)是将一对特异的抗原或抗体，其中一个固定于硝酸纤维素膜上，另一个用示踪物进行标记并制成结合垫，最后组装成试纸条；在试纸条的加样端，也就是样品垫的位置进行样本和稀释液的加样，样本由于层析力会沿着硝酸纤维素膜进行层析，若样本中有待检物，则待检物会和结合垫里面带了示踪物的抗原抗体结合，在经过硝酸纤维素膜上固定的抗原抗体处会被拦截，而显示一定的颜色，从而实现特异性的免疫诊断。免疫层析法，由于其简单、方便、快捷、便宜及不需要大型仪器和专业人员的操作而在疾病诊断、优生优育、食品检测、环境监测等领域得到了广泛的应用。免疫层析法中常用到的免疫层析试纸条主要包含五个部分：加样区(样品垫)、标记区(结合垫)、显示区(硝酸纤维素膜)、吸水区(吸水垫)以及背衬板。在传统的免疫层析试纸条中，吸水区是由AN3、AN5等滤纸常规未处理的吸水材料组成，为层析提供一定的层析力；产品内部的质控系统为C系统，在跑板时，一般通过质控线（C线）是否显色来确认跑板是否有效。
 技术问题
但是，传统C系统大多采用一些独立蛋白类系统，在样本层析过程中很容易和样本中的蛋白类物质发生交叉干扰，从而造成产品的特异性较差。因此，急需开发一种特异性高的免疫层析试纸条。
 技术解决方案
本发明提供了一种可变色指示的免疫层析试纸条及其使用方法，可以有效解决上述问题。
本发明是这样实现的：
一种可变色指示的免疫层析试纸条，包括底板和底板上沿层析方向依次连接的样品垫、结合垫、反应膜及吸水垫，所述反应膜上设置检测线，不设置质控线；所述吸水垫在检测样品液到达时变色。
一实施例中，所述吸水垫为经酸碱指示剂处理的吸水垫。
一实施例中，所述酸碱指示剂处理为将吸水垫浸泡在酸碱指示剂处理液中或将酸碱指示剂处理液涂抹在吸水垫上，一段时间后烘干备用。
一实施例中，还包括覆盖在所述的免疫层析试纸条上方的外壳，所述外壳上设有用于滴加检测样品液的加样区、用于观察或读取信号的检测区和用于判断跑板是否有效的指示区，所述加样区位于样品垫的上方，所述检测区位于检测线的上方，所述指示区位于吸水垫的上方。
一种上述的可变色指示的免疫层析试纸条的使用方法，包括以下步骤：
将检测样品液滴加在样品垫上，静置一段时间；
通过观察吸水垫是否变色来判断跑板是否有效，如果变色，则跑板有效，如果不变色，则跑板无效，需重新跑板；
通过观察检测线来读取检测结果。
一实施例中，所述检测样品液在滴加前加入碱性缓冲液或酸性缓冲液。
 有益效果
本发明的有益效果是：
本发明将变色的吸水垫应用于免疫层析试纸，不需采用传统C系统的独立蛋白类系统，避免在层析过程中发生交叉干扰，特异性高。
本发明将变色的吸水垫应用于免疫层析试纸，变色的吸水垫，相对于传统的蛋白类C系统，价格低廉，读取结果快捷而直观，不需要特殊仪器设备，也不需要专业人员操作，可以大量的节约物料成本和人力操作方面的成本。
 附图说明
为了更清楚地说明本发明实施方式的技术方案，下面将对实施方式中所需要使用的附图作简单地介绍，应当理解，以下附图仅示出了本发明的某些实施例，因此不应被看作是对范围的限定，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他相关的附图。
图1是本发明实施例提供的可变色指示的免疫层析试纸条的结构示意图。
图2是本发明实施例提供的可变色指示的免疫层析试纸条的外壳结构示意图。
附图标记：
样品垫1、结合垫2、反应膜3、检测线31、吸水垫4、底板5、加样区6、检测区7、指示区8。
 本发明的实施方式
为使本发明实施方式的目的、技术方案和优点更加清楚，下面将结合本发明实施方式中的附图，对本发明实施方式中的技术方案进行清楚、完整地描述，显然，所描述的实施方式是本发明一部分实施方式，而不是全部的实施方式。基于本发明中的实施方式，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方式，都属于本发明保护的范围。因此，以下对在附图中提供的本发明的实施方式的详细描述并非旨在限制要求保护的本发明的范围，而是仅仅表示本发明的选定实施方式。基于本发明中的实施方式，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方式，都属于本发明保护的范围。
在本发明的描述中，术语“第一”、“第二”仅用于描述目的，而不能理解为指示或暗示相对重要性或者隐含指明所指示的技术特征的数量。由此，限定有“第一”、“第二”的特征可以明示或者隐含地包括一个或者更多个该特征。在本发明的描述中，“多个”的含义是两个或两个以上，除非另有明确具体的限定。
实施例
如图1所示，本发明实施例提供一种可变色指示的免疫层析试纸条，包括底板5和底板5上沿层析方向依次连接的样品垫1、结合垫2、反应膜3及吸水垫4。所述反应膜3上设置检测线31，不设置质控线。所述吸水垫4在检测样品液到达时变色。如果检测完成后，所述吸水垫4变色，则说明用于检测样品液已经达到了吸水垫4，则必然已经经过了所述反应膜3上设置检测线31，可以判断跑板有效；反之，则跑板无效，需重新跑板检测。
一实施例中，所述吸水垫4为经酸碱指示剂处理的吸水垫。检测样品液一般呈酸性或碱性，当检测样品液到达所述吸水垫时，所述吸水垫上的酸碱指示剂就会变色，表明跑板有效。
一实施例中，所述酸碱指示剂处理为将吸水垫浸泡在酸碱指示剂处理液中或将酸碱指示剂处理液涂抹在吸水垫上，一段时间后烘干备用。
一实施例中，所述酸碱指示剂处理液的配方为53wt%的乙醇、0.0047wt%的甲基红、0.10wt%的溴百里酚蓝、0.45wt%的20wt%Tween20、18wt%的5wt%PVA、19wt%的5wt%PVP 溶液。
一实施例中，所述可变色指示的免疫层析试纸条还包括覆盖在所述的免疫层析试纸条上方的外壳，所述外壳上设有用于滴加检测样品液的加样区6、用于观察或读取信号的检测区7和用于判断跑板是否有效的指示区8，所述加样区6位于样品垫1的上方，所述检测区7位于检测线31的上方，所述指示区8位于吸水垫4的上方。
一实施例中，所述样品垫1、结合垫2、反应膜3、吸水垫4的长度比为2：1：4：1；此长度比例便于样品加到所述样品垫1上，同时反应膜3足够长便于样品液停留在反应膜3上足够时间以完成检测反应。所述样品垫1与结合垫2在连接处重叠，且所述样品垫1重叠在结合垫2的上方，重叠部分的长度为结合垫2的长度的1/4-1/2，便于样品液顺利地从所述样品垫1流到结合垫2上。所述结合垫2与反应膜3在连接处重叠，且所述结合垫2重叠在反应膜3的上方，重叠部分的长度为反应膜3的长度的1/8-1/4；便于样品液顺利地从所述结合垫2流到反应膜3上。所述反应膜3与吸水垫4在连接处重叠，且吸水垫4重叠在反应膜3的上方，重叠部分的长度为吸水垫4的长度的1/5-1/3，便于样品液顺利地从所述反应膜3吸附到吸水垫4上。检测线31位于反应膜3的1/2处。如此的尺寸设置，便于样品液层析沿着所述样品垫1、结合垫2、反应膜3、吸水垫4的方向流动，同时样品液停留在反应膜3上足够时间以完成检测反应，提高检测的灵敏度和效率。
一实施例中，所述样品垫1为玻璃纤维膜或聚酯纤维膜。
一实施例中，所述结合垫2为玻璃纤维膜或聚酯纤维膜。
一实施例中，所述反应膜3为硝酸纤维素膜。
一实施例中，所述吸水垫4为滤纸（如AN3、AN5)
一实施例中，所述底板5为PVC底板或PS底板。
一实施例中，所述检测样品液为血清、全血、尿液、组织液、咽拭子、鼻拭子、口咽拭子、肛拭子或排泄物等的一种或多种。
本发明实施例还提供一种上述的可变色指示的免疫层析试纸条的使用方法，包括以下步骤：
将检测样品液滴加在样品垫1上，滴加的量为1-3滴，静置一段时间，通常为10-20min，检测样品液沿着样品垫1、结合垫2、反应膜3及吸水垫4层析，层析到吸水垫上时检测完成；
通过观察吸水垫4是否变色来判断跑板是否有效，如果变色，则跑板有效，如果不变色，则跑板无效，需重新跑板；
通过观察检测线31来读取检测结果。
一实施例中，所述检测样品液在滴加前加入碱性缓冲液或酸性缓冲液。碱性缓冲液或酸性缓冲液的pH可以根据酸碱指示剂处理液的类型进行调整。进一步优选为加入碱性缓冲液。
试验例1
在登革抗原检测项目中，测试实施例1的可变色指示的免疫层析试纸条的检测特异性。测试方法为采用很多阴性样本来进行检测实验，看是否会出现假阳样本，从而来统计特异性。传统蛋白类类C线系统的免疫层析试纸条作为对照组，实施例1的可变色指示的免疫层析试纸条作为实验组。两组均做血清200例，血浆200例，全血200例。统计实验结果如表1所示。
表1
组别 特异性 样本类型 血清 血浆 全血
对照组 96.5% 97% 96.5%
实验组 98% 98.5% 99.5%
由表1可知，实施例1的可变色指示的免疫层析试纸条的检测特异性要高于传统蛋白类类C线系统的免疫层析试纸条。分析原因认为，本发明实施例1的可变色指示的免疫层析试纸条，不设置质控线，不需采用传统C系统的独立蛋白类系统，避免在层析过程中发生蛋白的交叉干扰，提高了特异性。
试验例2
在甲型/乙型流感病毒抗原检测项目中，测试实施例1的可变色指示的免疫层析试纸条的检测特异性。测试方法为采用很多阴性样本来进行检测实验，看是否会出现假阳样本，从而来统计特异性。传统蛋白类类C线系统的免疫层析试纸条作为对照组，实施例1的可变色指示的免疫层析试纸条作为实验组。两组均做阴性血清样本500例。统计实验结果如表2所示。
表2
  组别 组别 特异性 特异性
对照组 98.8%
实验组 99.6%
由表2可知，实施例1的可变色指示的免疫层析试纸条的检测特异性要高于传统蛋白类类C线系统的免疫层析试纸条。分析原因认为，本发明实施例1的可变色指示的免疫层析试纸条，不设置质控线，不需采用传统C系统的独立蛋白类系统，避免在层析过程中发生蛋白的交叉干扰，提高了特异性。
试验例3
在甲型/乙型流感病毒抗原检测项目中，传统蛋白类类C线系统的免疫层析试纸条作为对照组，实施例1的可变色指示的免疫层析试纸条作为实验组，比较两者的背景完全跑干净时间。背景完全跑干净判断标准：视窗无明显的红色背景。统计结果表3所示。
表3
  组别 组别 背景干净时间（）（）（S） 样本本 1 2 3 4 5 6 7 8 9 10 11 12 13 14 15
对照组 90s 89s 93s 88s 90s 91s 95s 87s 80s 85s 89s 91s 92s 91s 89s
实验组 80s 79s 78s 77s 78s 79s 76s 79s 75s 76s 75s 73s 75s 74s 76s
由表3可知，实施例1的可变色指示的免疫层析试纸条的背景干净时间更短，说明其检测效率更高。分析原因认为，本发明的实施例1的可变色指示的免疫层析试纸条不设置质控线，减少了一道反应程序，检测样品溶液在反应膜上停留的时间更短，从而缩短了了整个背景干净时间，提高了检测效率。
以上所述仅为本发明的优选实施方式而已，并不用于限制本发明，对于本领域的技术人员来说，本发明可以有各种更改和变化。凡在本发明的精神和原则之内，所作的任何修改、等同替换、改进等，均应包含在本发明的保护范围之内。

Claims (6)

  1. 一种可变色指示的免疫层析试纸条，包括底板和底板上沿层析方向依次连接的样品垫、结合垫、反应膜及吸水垫，其特征在于，所述反应膜上设置检测线，不设置质控线；所述吸水垫在检测样品液到达时变色；所述吸水垫为经酸碱指示剂处理的吸水垫；所述样品垫与结合垫在连接处重叠，且所述样品垫重叠在结合垫的上方；所述结合垫与反应膜在连接处重叠，且所述结合垫重叠在反应膜的上方；所述反应膜与吸水垫在连接处重叠，且吸水垫4重叠在反应膜3的上方。
  2. 根据权利要求1所述的可变色指示的免疫层析试纸条，其特征在于，所述酸碱指示剂处理为将吸水垫浸泡在酸碱指示剂处理液中或将酸碱指示剂处理液涂抹在吸水垫上，一段时间后烘干备用。
  3. 根据权利要求1或2所述的可变色指示的免疫层析试纸条，其特征在于，所述酸碱指示剂的配方为53wt%的乙醇、0.0047wt%的甲基红、0.10wt%的溴百里酚蓝、0.45wt%的20wt%Tween20、18wt%的5wt%PVA、19wt%的5wt%PVP 溶液。
  4. 根据权利要求1所述的可变色指示的免疫层析试纸条，其特征在于，还包括覆盖在所述的免疫层析试纸条上方的外壳，所述外壳上设有用于滴加检测样品液的加样区、用于观察或读取信号的检测区和用于判断跑板是否有效的指示区，所述加样区位于样品垫的上方，所述检测区位于检测线的上方，所述指示区位于吸水垫的上方。
  5. 一种权利要求1至4所述的可变色指示的免疫层析试纸条的使用方法，其特征在于，包括以下步骤：
    将检测样品液滴加在样品垫上，静置一段时间；
    通过观察吸水垫是否变色来判断跑板是否有效，如果变色，则跑板有效，如果不变色，则跑板无效，需重新跑板；
    通过观察检测线来读取检测结果。
  6. 根据权利要求5所述的方法，其特征在于，所述检测样品液在滴加前加入碱性缓冲液或酸性缓冲液。
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