Self-amplification precisely-controlled indirect competitive immunochromatography test strip and application thereof

An immunochromatographic detection and test strip technology, applied in analytical materials, biological testing, biological material analysis, etc., can solve the problems of unsuccessful establishment of immunochromatographic test strips, low efficiency of artificial antigen interception antibodies, and no signal amplification effect. , to achieve the effect of increasing the sensitivity of the test paper, high sensitivity and display image

Pending Publication Date: 2021-06-25
HENAN ACAD OF AGRI SCI
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] In the field of food safety, small molecule antigen immunochromatography technology based on the principle of competition is the most widely used. However, the existing detection mode often encounters immune reagents such as antigens and antibodies. ELISA detection methods can be successfully established, but immunochromatographic test strips cannot be successfully established; And in many applications the sensitivity of the test strip still needs to be improved
The main reasons are as follows: 1. The antibody folded during the labeling process, and an excessive amount of antibody was added to stabilize the labeling material; 2. The efficiency of the artificial antigen intercepting antibody was low, and an excessive amount of labeled antibody had to be added in order to improve color development. Improve color development; 3. There is no signal amplification when colloidal gold is used to label antibodies. Fluorescent materials such as quantum dots achieve signal amplification while requiring detection instruments to judge the results, which complicates detection.
The existing small molecule immunochromatography detection mode cannot solve this problem, so a simple and sensitive test strip detection mode is urgently needed to provide a better tool for the development of immunochromatography test strips

Method used

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  • Self-amplification precisely-controlled indirect competitive immunochromatography test strip and application thereof
  • Self-amplification precisely-controlled indirect competitive immunochromatography test strip and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0031] A "self-amplification" and "precise control" 2,4-D indirect competitive immunochromatography test strip, which consists of two parts: a test paper and a sample cup; the test paper includes a support layer and an adsorption layer fixed on the support layer and a protective layer; the adsorption layer is followed by the sample pad, the binding pad, the cellulose film layer and the water-absorbing material layer at the handle end from the test end; the protective layer is fixed on the sample pad, the binding pad and the water-absorbing material layer; the cellulose film The detection line and the quality control line are arranged in turn on the layer; the binding pad is made of glass fiber cotton adsorbed with nanomaterial-labeled antigen; the sample cup is a container for diluting samples, and the weight of gold-labeled protein is pre-fixed in the sample cup. 2,4-D monoclonal antibody diluted in suspension.

[0032] The nanomaterial labeled antigen is a 2,4-D artificial ant...

Embodiment 2

[0038] A method for preparing a "self-amplifying" and "precisely controlled" 2,4-D indirect competitive immunochromatography test strip, comprising the following steps:

[0039] 1. Preparation of 2,4-D artificial antigen

[0040] Weigh 100 mg of 2,4-D and add 200 μL of methanol to dissolve, then add 7 mL of PBS and the solution becomes turbid, then add 1 mol / L NaOH solution drop by drop until the solution is clear, then adjust the pH value of the solution to neutral with 0.1M HCl, which is A Solution; Weigh 15mg bovine serum albumin (BSA) and dissolve it in 300μL PBS. After fully dissolving, this is solution B; slowly add solution A to solution B under stirring at 4°C, and then add 100mg carbodiimide (EDC ), reacted at 4°C under stirring for 12h, took out the reaction product, and dialyzed it with PBS for 72h to obtain 2,4-D-BSA, which was frozen at -20°C for future use.

[0041] 2. Preparation of 2,4-D monoclonal antibody

[0042] Use the prepared 2,4-D-BSA to immunize 6-8 ...

Embodiment 3

[0060] A "self-amplifying" and "precisely controlled" gentamicin indirect competitive immunochromatography test strip consists of two parts: a test paper and a sample cup; the test paper includes a support layer, an adsorption layer fixed on the support layer and Protective layer; the adsorption layer is followed by the sample pad, the binding pad, the cellulose film layer and the water-absorbing material layer at the handle end from the test end; the protective layer is fixed on the sample pad, the binding pad and the water-absorbing material layer; the cellulose film layer The test line and the quality control line are arranged in sequence on the top; the binding pad is made of glass fiber cotton adsorbed with nanomaterial-labeled antigen; the sample cup is a container for diluting samples, and gold-labeled protein suspension is pre-immobilized on it. Diluted gentamicin monoclonal antibody.

[0061]The nanomaterial labeled antigen is a gentamicin artificial antigen labeled w...

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Abstract

The invention relates to a 'self-amplification' and 'precise control' indirect competitive immunochromatography detection test strip which consists of two parts, namely test paper and a sample cup. The test paper comprises a supporting layer, an adsorption layer fixed on the supporting layer and a protective layer. The adsorption layer sequentially comprises a sample pad, a combination pad, a cellulose membrane layer and a water absorption material layer at a handle end from a test end. A detection line and a quality control line are sequentially arranged on the cellulose membrane layer. The conjugate pad is made of glass fiber cotton adsorbed with a nano material labeled antigen. The sample cup is a container for diluting a sample, and a target monoclonal antibody is pre-fixed in the sample cup. According to the test strip, the antibody is independently dried in the sample cup, so that folding and damage in the antibody labeling process are avoided, and the antibody dosage can be accurately controlled; and nano-particle labeled artificial antigens are used as a probe, and each antibody intercepted on a detection line can be accurately combined with two labeled antigens, so that signal self-amplification is realized, and the sensitivity of the test paper is improved.

Description

technical field [0001] The invention relates to an immunochromatographic detection method, in particular to an indirect competitive immunochromatographic detection test strip with "self-amplification" and "precise control" for hapten detection and its application. Background technique [0002] Immunochromatography is a rapid immunoassay technology developed in recent years. Its basic principle is the interaction between antibodies and antigens. It uses nitrocellulose membrane as a carrier to label antigens or antibodies as a tracer. A chromatographic detection technique. Compared with traditional immunoassay methods, immunochromatography has the advantages of strong specificity, accurate detection results, simple equipment operation, rapid measurement, low cost, no need for skilled technicians or expensive equipment, etc., fully meeting the requirements of instant detection . At present, immunochromatography technology has been widely used in medicine, animal husbandry, ag...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/558G01N33/577G01N33/58G01N33/53G01N33/532G01N33/531
CPCG01N33/5308G01N33/531G01N33/532G01N33/558G01N33/577G01N33/587G01N33/588G01N33/9446G01N2430/20
Inventor 孙亚宁杨苏珍郭振华金前跃杨继飞胡骁飞邓瑞广张改平
Owner HENAN ACAD OF AGRI SCI
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