CN115327111A - Immunochromatography test strip with variable color indication and use method thereof - Google Patents

Immunochromatography test strip with variable color indication and use method thereof Download PDF

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Publication number
CN115327111A
CN115327111A CN202210905760.1A CN202210905760A CN115327111A CN 115327111 A CN115327111 A CN 115327111A CN 202210905760 A CN202210905760 A CN 202210905760A CN 115327111 A CN115327111 A CN 115327111A
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pad
test strip
detection
sample
immunochromatographic test
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陈亚茹
赵弟萍
倪建良
杨姗姗
张国锋
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Hangzhou Baotai Biotechnology Co ltd
Xiamen Baotai Biotechnology Co ltd
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Hangzhou Baotai Biotechnology Co ltd
Xiamen Baotai Biotechnology Co ltd
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Priority to CN202210905760.1A priority Critical patent/CN115327111A/en
Publication of CN115327111A publication Critical patent/CN115327111A/en
Priority to PCT/CN2023/085581 priority patent/WO2024021664A1/en
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    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N33/00Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
    • G01N33/48Biological material, e.g. blood, urine; Haemocytometers
    • G01N33/50Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
    • G01N33/53Immunoassay; Biospecific binding assay; Materials therefor
    • G01N33/558Immunoassay; Biospecific binding assay; Materials therefor using diffusion or migration of antigen or antibody
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/01Arrangements or apparatus for facilitating the optical investigation
    • GPHYSICS
    • G01MEASURING; TESTING
    • G01NINVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/75Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
    • G01N21/77Systems 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
    • G01N21/78Systems 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

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  • Urology & Nephrology (AREA)
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  • Investigating Or Analysing Materials By The Use Of Chemical Reactions (AREA)

Abstract

The invention provides an immunochromatographic test strip with variable color indication and a use method thereof, and the immunochromatographic test strip comprises a bottom plate and a sample pad, a combination pad, a reaction membrane and a water absorption pad which are sequentially connected on the bottom plate along a chromatography direction, and is characterized in that the reaction membrane is provided with a detection line and is not provided with a quality control line; the absorbent pad changes color when the detection sample fluid arrives. The immunochromatographic test strip with the color-variable indication has high detection specificity and low cost.

Description

Immunochromatography test strip with variable color indication and use method thereof
Technical Field
The invention relates to an immunochromatographic test strip with variable color indication and a use method thereof, belonging to the technical field of immunochromatographic.
Background
The immunochromatography (immunochromatography) is to fix a pair of specific antigens or antibodies, wherein one of the specific antigens or antibodies is fixed on a nitrocellulose membrane, and the other specific antigen or antibody is marked by a tracer and made into a binding pad, and finally assembled into a test strip; the sample and the diluent are added at the sample adding end of the test strip, namely the position of the sample pad, the sample can carry out chromatography along the nitrocellulose membrane due to the chromatographic force, if an object to be detected is to be detected in the sample, the object to be detected can be combined with the antigen-antibody which is provided with the tracer in the combination pad, the fixed antigen-antibody position on the nitrocellulose membrane can be intercepted, and a certain color is displayed, so that the specific immunodiagnosis is realized. Immunochromatography is widely used in the fields of disease diagnosis, prenatal and postnatal care, food detection, environmental monitoring and the like because it is simple, convenient, rapid, inexpensive and does not require large-scale instruments and professional operations. An immunochromatography test strip commonly used in immunochromatography mainly comprises five parts: a sample adding area (sample pad), a labeling area (combination pad), a display area (nitrocellulose membrane), a water absorbing area (water absorbing pad) and a backing plate. In the traditional immunochromatographic test strip, a water absorption area is formed by conventional untreated water absorption materials of filter paper such as AN3, AN5 and the like, and a certain chromatographic power is provided for chromatography; the quality control system in the product is a C system, and when the running board is run, whether the running board is effective or not is generally determined by whether a quality control line (C line) is developed or not. However, most of the conventional C systems adopt some independent protein systems, which are easily cross-interfered with protein substances in a sample during sample chromatography, thereby resulting in poor product specificity. Therefore, it is urgently needed to develop an immunochromatographic test strip with high specificity.
Disclosure of Invention
The invention provides an immunochromatographic test strip with a color-variable indication function and a use method thereof, which can effectively solve the problems.
The invention is realized by the following steps:
an immunochromatography test strip with color-variable indication comprises a bottom plate and a sample pad, a combination pad, a reaction membrane and a water absorption pad which are sequentially connected on the bottom plate along a chromatography direction, wherein the reaction membrane is provided with a detection line and is not provided with a quality control line; the absorbent pad changes color when the detection sample liquid arrives.
In one embodiment, the absorbent pad is treated with an acid-base indicator.
In one embodiment, the acid-base indicator treatment is to soak the water absorbent pad in an acid-base indicator treatment solution or to smear the acid-base indicator treatment solution on the water absorbent pad, and then dry the water absorbent pad for later use after a period of time.
In one embodiment, still including covering the shell of immunochromatographic test strip top, be equipped with the area of application of sample that is used for dropwise add the test sample liquid, be used for observing or the detection zone of reading signal and be used for judging whether effectual instruction zone of race plate on the shell, the area of application of sample is located the top of sample pad, the detection zone is located the top of detection line, the instruction zone is located the top that absorbs water and fills up.
The use method of the immunochromatographic test strip with the color-variable indication function comprises the following steps:
dropwise adding a detection sample solution on a sample pad, and standing for a period of time;
judging whether the running board is effective or not by observing whether the water absorption pad changes color or not, if so, judging that the running board is effective, and if not, judging that the running board is ineffective and needing to run again;
the detection result is read by observing the detection line.
In one embodiment, the detection sample solution is added with an alkaline buffer solution or an acidic buffer solution before being added dropwise.
The invention has the beneficial effects that:
the color-changing absorbent pad is applied to the immunochromatographic test paper, an independent protein system of the traditional C system is not needed, cross interference in the chromatography process is avoided, and the specificity is high.
Compared with the traditional protein C system, the color-changing absorbent pad has the advantages of low price, quick and visual reading result, no need of special instruments and equipment, no need of professional operation, and capability of greatly saving material cost and cost in the aspect of manual operation.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are required to be used in the embodiments will be briefly described below, it should be understood that the following drawings only illustrate some embodiments of the present invention and therefore should not be considered as limiting the scope, and for those skilled in the art, other related drawings can be obtained according to the drawings without inventive efforts.
Fig. 1 is a schematic structural diagram of an immunochromatographic test strip with a variable color indicator provided in an embodiment of the present invention.
Fig. 2 is a schematic structural diagram of a housing of the immunochromatographic test strip with a variable color indicator provided in the embodiment of the present invention.
Reference numerals:
sample pad 1, combination pad 2, reaction membrane 3, detection line 31, absorbent pad 4, bottom plate 5, application of sample district 6, detection zone 7, instruction district 8.
Detailed Description
In order to make the objects, technical solutions and advantages of the embodiments of the present invention more apparent, the technical solutions of the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings of the embodiments of the present invention, and it is obvious that the described embodiments are some, but not all embodiments of the present invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention. Thus, the following detailed description of the embodiments of the present invention, as presented in the figures, is not intended to limit the scope of the invention, as claimed, but is merely representative of selected embodiments of the invention. All other embodiments, which can be obtained by a person skilled in the art without any inventive step based on the embodiments of the present invention, are within the scope of the present invention.
In the description of the present invention, the terms "first" and "second" are used for descriptive purposes only and are not to be construed as indicating or implying relative importance or implying any number of technical features indicated. Thus, a feature defined as "first" or "second" may explicitly or implicitly include one or more of that feature. In the description of the present invention, "a plurality" means two or more unless specifically defined otherwise.
Examples
As shown in fig. 1, an embodiment of the present invention provides a color-changeable indicator immunochromatographic test strip, which includes a bottom plate 5, and a sample pad 1, a conjugate pad 2, a reaction membrane 3, and a water absorption pad 4, which are sequentially connected to the bottom plate 5 along a chromatography direction. And the reaction membrane 3 is provided with a detection line 31 and is not provided with a quality control line. The absorbent pad 4 changes color when the detection sample liquid arrives. If the absorbent pad 4 changes color after the detection is finished, the detection line 31 is set on the reaction membrane 3 inevitably after the sample liquid reaches the absorbent pad 4, so that the running board can be judged to be effective; otherwise, the running board is invalid, and the running board detection is needed again.
In one embodiment, the absorbent pad 4 is an absorbent pad treated with an acid-base indicator. The detection sample liquid is generally acidic or alkaline, and when the detection sample liquid reaches the water absorption pad, the acid-base indicator on the water absorption pad changes color, so that the running board is effective.
In one embodiment, the acid-base indicator treatment is to soak the water absorbent pad in an acid-base indicator treatment solution or to smear the acid-base indicator treatment solution on the water absorbent pad, and then dry the water absorbent pad for later use after a period of time.
In one embodiment, the acid-base indicator treatment fluid is formulated with 53wt% ethanol, 0.0047wt% methyl red, 0.10wt% bromothymol blue, 0.45wt% 20wt% tween20, 18wt% 5wt% pva, 19wt% 5wt% pvp solution.
In one embodiment, the immunochromatographic test strip with the color-changeable indication further comprises a shell covering the immunochromatographic test strip, wherein the shell is provided with a sample adding region 6 for dropwise adding a detection sample solution, a detection region 7 for observing or reading a signal and an indication region 8 for judging whether a running plate is effective or not, the sample adding region 6 is positioned above the sample pad 1, the detection region 7 is positioned above the detection line 31, and the indication region 8 is positioned above the water absorption pad 4.
In one embodiment, the sample pad 1 overlaps the conjugate pad 2 at the junction, and the sample pad 1 overlaps the conjugate pad 2; the bonding pad 2 overlaps the reaction membrane 3 at the joint, and the bonding pad 2 overlaps the reaction membrane 3; the reaction membrane 3 is overlapped with the absorbent pad 4 at the joint, and the absorbent pad 4 is overlapped above the reaction membrane 3. Thus, the sample solution is conveniently chromatographed and stays on the reaction membrane 3 for a sufficient time to complete the detection reaction.
In one embodiment, the sample pad 1 is a glass fiber film or a polyester fiber film.
In one embodiment, the bonding pad 2 is a glass fiber film or a polyester fiber film.
In one embodiment, the reaction membrane 3 is a nitrocellulose membrane.
In one embodiment, the absorbent pad 4 is filter paper (e.g., AN3, AN 5)
In one embodiment, the bottom board 5 is a PVC bottom board or a PS bottom board.
In one embodiment, the sample fluid to be tested is one or more of serum, whole blood, urine, tissue fluid, throat swab, nasal swab, oropharynx swab, anal swab, excrement, and the like.
The embodiment of the invention also provides a use method of the immunochromatographic test strip with the color-variable indication function, which comprises the following steps:
dropwise adding a detection sample solution on the sample pad 1, standing for a period of time, usually 10-20min, carrying out chromatography on the detection sample solution along the sample pad 1, the combination pad 2, the reaction membrane 3 and the water absorption pad 4, and completing detection when the detection sample solution is chromatographed on the water absorption pad;
whether the running board is effective or not is judged by observing whether the water absorption pad 4 changes color, if the color is changed, the running board is effective, and if the color is not changed, the running board is ineffective, and the running board needs to be run again;
the detection result is read by observing the detection line 31.
In one embodiment, the detection sample solution is added with an alkaline buffer solution or an acidic buffer solution before being added dropwise. The pH of the alkaline buffer or acidic buffer may be adjusted depending on the type of acid-base indicator treatment fluid. Further preferably, an alkaline buffer is added.
Test example 1
The immunochromatographic test strip of variable color indication of example 1 was tested for detection specificity in the dengue antigen detection project. The test method is to adopt a plurality of negative samples to carry out detection experiments to see whether false positive samples appear or not, thereby counting the specificity. The immunochromatographic test strip of the traditional protein C-line system serves as a control group, and the immunochromatographic test strip with variable color indication in example 1 serves as an experimental group. 200 sera, 200 plasma and 200 whole blood were prepared for both groups. The statistical test results are shown in table 1.
TABLE 1
Figure BDA0003772372620000061
As can be seen from table 1, the detection specificity of the immunochromatographic test strip indicated by variable color in example 1 is higher than that of the immunochromatographic test strip of the conventional protein C-line system. For analysis reasons, the immunochromatographic test strip with variable color indication in embodiment 1 of the present invention does not need to set a quality control line, and does not need to use an independent protein system of the conventional C system, thereby avoiding cross interference of proteins during chromatography and improving specificity.
Test example 2
The immunochromatographic test strip of the variable color indicator of example 1 was tested for detection specificity in the influenza a/b virus antigen detection project. The test method is to adopt a plurality of negative samples to carry out detection experiments to see whether false positive samples appear or not, thereby counting the specificity. The immunochromatographic test strip of the traditional protein C-line system serves as a control group, and the immunochromatographic test strip with variable color indication in example 1 serves as an experimental group. Both groups were used as 500 negative serum samples. The statistical test results are shown in table 2.
TABLE 2
Figure BDA0003772372620000071
As can be seen from table 2, the detection specificity of the immunochromatographic test strip indicated by variable color in example 1 is higher than that of the immunochromatographic test strip of the conventional protein C-line system. For analysis reasons, the immunochromatographic test strip with variable color indication in embodiment 1 of the present invention does not need to set a quality control line, and does not need to use an independent protein system of the conventional C system, thereby avoiding cross interference of proteins during chromatography and improving specificity.
Test example 3
In the influenza a/b virus antigen detection project, the immunochromatographic test strip of the conventional protein type C line system is used as a control group, the immunochromatographic test strip indicated by variable color in example 1 is used as an experimental group, and the background of the two is compared with the total clean time. And (3) completely running the background to a clean judgment standard: the window had no visible red background. Statistical results are shown in table 3.
TABLE 3
Figure BDA0003772372620000081
As can be seen from table 3, the immunochromatographic test strip of example 1 with variable color indication has a shorter background cleaning time, indicating that the detection efficiency is higher. The reason for analysis is that the immunochromatographic test strip with variable color indication in example 1 of the present invention does not have a quality control line, one reaction procedure is reduced, and the stay time of the detection sample solution on the reaction membrane is shorter, thereby shortening the whole background clean time and improving the detection efficiency.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, and various modifications and changes may be made by those skilled in the art. Any modification, equivalent replacement, or improvement made within the spirit and principle of the present invention should be included in the protection scope of the present invention.

Claims (6)

1. An immunochromatographic test strip with a color-variable indication function comprises a base plate and a sample pad, a combination pad, a reaction membrane and a water absorption pad which are sequentially connected with the base plate along a chromatography direction, and is characterized in that the reaction membrane is provided with a detection line and is not provided with a quality control line; the absorbent pad changes color when the detection sample fluid arrives.
2. The immunochromatographic test strip for variable color indication according to claim 1, wherein the absorbent pad is an absorbent pad treated with an acid-base indicator.
3. The immunochromatographic test strip with variable color indication according to claim 1, wherein the acid-base indicator treatment is to soak a water absorption pad in an acid-base indicator treatment solution or to coat the acid-base indicator treatment solution on the water absorption pad, and after a period of time, the water absorption pad is dried for standby.
4. The immunochromatographic test strip with a variable color indicator according to claim 1, further comprising a housing covering the immunochromatographic test strip, wherein the housing is provided with a sample adding region for dropwise adding a detection sample solution, a detection region for observing or reading a signal, and an indication region for judging whether a running board is effective, the sample adding region is located above the sample pad, the detection region is located above the detection line, and the indication region is located above the absorbent pad.
5. The method for using the immunochromatographic test strip with variable color indication according to claims 1 to 4, which is characterized by comprising the following steps:
dropwise adding a detection sample solution on a sample pad, and standing for a period of time;
judging whether the running board is effective or not by observing whether the water absorption pad changes color or not, if so, judging that the running board is effective, and if not, judging that the running board is ineffective and needing to run again;
the detection result is read by observing the detection line.
6. The method of claim 5, wherein the test sample solution is added with an alkaline buffer or an acidic buffer before the dropwise addition.
CN202210905760.1A 2022-07-29 2022-07-29 Immunochromatography test strip with variable color indication and use method thereof Pending CN115327111A (en)

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PCT/CN2023/085581 WO2024021664A1 (en) 2022-07-29 2023-03-31 Immunochromatographic test strip capable of changing color for indication and use method thereof

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024021664A1 (en) * 2022-07-29 2024-02-01 厦门宝太生物科技股份有限公司 Immunochromatographic test strip capable of changing color for indication and use method thereof

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CN115327111A (en) * 2022-07-29 2022-11-11 厦门宝太生物科技股份有限公司 Immunochromatography test strip with variable color indication and use method thereof

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WO1992019764A1 (en) * 1991-05-08 1992-11-12 Baxter Diagnostics Inc. Method and apparatus to detect bacterial contamination of transfusable blood
US5415994A (en) * 1993-08-02 1995-05-16 Quidel Corporation Lateral flow medical diagnostic assay device with sample extraction means
WO1998039657A1 (en) * 1997-03-06 1998-09-11 Quidel Corporation Quantitative lateral flow assays and devices
US6413473B1 (en) * 1999-02-05 2002-07-02 Taylor Technologies, Inc. Multicomponent test systems useful in analyzing liquid samples, and uses therefor
JP2011017689A (en) * 2009-06-12 2011-01-27 Rohto Pharmaceutical Co Ltd Urine examination implement and vessel
CN106198527A (en) * 2016-07-13 2016-12-07 青岛汉唐生物科技有限公司 A kind of ascorbic acid interference multi-term urine analysis test paper
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* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2024021664A1 (en) * 2022-07-29 2024-02-01 厦门宝太生物科技股份有限公司 Immunochromatographic test strip capable of changing color for indication and use method thereof

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Application publication date: 20221111