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Double-anti-sandwich colloidal gold detection kit, and preparation method and application thereof

A double-antibody sandwich and colloidal gold technology, applied in the field of rapid diagnosis and detection of epidemic diseases, can solve problems such as unusable, lack of double-antibody sandwich test strip sample processing kits, and reduced sensitivity

Active Publication Date: 2019-10-11
LUOYANG PULIKE WANTAI BIOTECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0003] However, in the existing technology and trial and error, it was found that the rapid detection product involved in the principle of double antibody (double monoclonal antibody) sandwich, after confirming that two kinds of antibodies can be used to detect antigens in the double antibody sandwich, one of them was found when studying the collocation mode The monoclonal antibody can only be fixed for labeling antibodies, and the other monoclonal antibody can only be used as an antibody for fixing on the detection line. If the matching mode of the two antibodies is changed, that is, the positions of the two monoclonal antibodies in the product are exchanged, then Can cause non-specific reactions or reduce sensitivity
[0004] In addition, the sample treatment solutions in existing colloidal gold products contain different components. When dealing with non-targeted animal disease antigen samples, the antigens are not fully lysed, and the sample treatment solutions of different kits are only for speci

Method used

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  • Double-anti-sandwich colloidal gold detection kit, and preparation method and application thereof
  • Double-anti-sandwich colloidal gold detection kit, and preparation method and application thereof
  • Double-anti-sandwich colloidal gold detection kit, and preparation method and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0084] Example 1 Analysis of Monoclonal Antibody Situation Used in Existing Products

[0085] Based on the principle of double-antibody sandwich, the technical team of the present invention and other R&D teams have developed a series of products, which are briefly as follows:

[0086] In Chinese patent CN105717293A, when the porcine circovirus type 2 monoclonal antibody 2F8 (abbreviated as PCV2-2F8) and 3G12 (abbreviated as PCV2-3G12) were paired for antibody activity detection, it was found that only when the coated antibody was PCV2-2F8 and the labeled antibody PCV2-3G12 Only the prepared enzyme immunochromatography test strips can be used. If the two kinds of antibodies are exchanged, that is, when the coated antibody is PCV2-3G12 and the labeled antibody PCV2-2F8, there will be a non-specific reaction. The same problem occurred when 2 kinds of monoclonal antibodies were assembled into colloidal gold detection test strips.

[0087] In the Chinese patent CN105461805A, porci...

Embodiment 2

[0094] The preparation of embodiment 2 kit

[0095] For better evaluation of chromatography test strips such as canine parvovirus colloidal gold detection test strips, the present invention first prepares the test strips with the immobilized antibody as 10H4 and the labeled antibody as 10B11 (corresponding to the part of Example 2.1), and Canine parvovirus CVCCAV298 strain (purchased from China Veterinary Microorganism Culture Collection Management Center), S2 strain, S0425 strain, S0304 strain of different genotypes of canine parvovirus epidemic strain CPV (see Preparation and application of two monoclonal antibodies against canine parvovirus vaccine and field strains, Journal of Vaccines and Immunology, 2017, 3 (1): 001-004), and 20 positive feces and 20 negative feces (respectively identified as positive and negative by canine parvovirus PCR) as sample discs, and the test paper Articles are fully regulated.

[0096] 2.1 Preparation of test strips

[0097] 2.1.1 Preparatio...

Embodiment 3

[0187] Preparation of other colloidal gold detection test strips of embodiment 3

[0188] In order to further evaluate whether each condition optimized in Example 2 can meet the application in the diagnosis and detection of animal diseases such as pigs, poultry, cats, and other antigens in dogs, porcine circovirus type 2, porcine epidemic diarrhea virus, and porcine infectivity Gastroenteritis virus, type A influenza virus, canine distemper virus, canine adenovirus, etc. were debugged and evaluated one by one. The results: (1) Sensitivity. The sensitivity of the kit prepared in the original matching mode was better than that of the commercially available or before debugging (higher by 2 to 20 times), the sensitivity of the kit prepared by the matching mode after the exchange of positions is equivalent to or better than that of the original matching mode before debugging; (2) the specificity is good, and no non-specific phenomenon occurs; (3) ) repeatability, all good; (4) shel...

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Abstract

The present invention provides a double-anti-sandwich colloidal gold detection kit. A gold-labeled antibody blocking solution, a gold-labeled antibody dilution solution and an immobilized antibody dilution solution with specific components are used in the kit. According to the kit provided by the present invention, by using three reagents with specific components, the gold-labeled antibody and theimmobilized antibody in the original colloidal gold detection can be interchangeable; and when the three reagents are used, the detection sensitivity is increased or equivalent to the original colloidal gold detection result.

Description

technical field [0001] The invention relates to a test kit for a double-antibody sandwich colloidal gold detection test strip, a preparation method and application thereof, and belongs to the field of rapid diagnosis and detection of epidemic diseases. Background technique [0002] At present, in the field of rapid diagnosis and detection of epidemic diseases, it is common to use double-antibody sandwiches to detect antigens, such as Lu Zihua et al. Preparation of paper strips. Colloidal gold detection test strips based on the monoclonal antibody-polyclonal antibody sandwich principle reported in Livestock Production and Poultry Disease Control, 597) to detect goose paramyxovirus antigens; and Dong-Jun An et al. ( See Dong-Jun An et al. Animmunochromatography assay for rapid antemortem diagnosis of dogs suspected to have canine distemper. Journal of Virological Methods, 2008, 147:244-249) reported in the colloidal gold based on the monoclonal antibody-monoclonal antibody san...

Claims

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Application Information

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IPC IPC(8): G01N33/577G01N33/532
CPCG01N33/577G01N33/532Y02A50/30
Inventor 田克恭王莹
Owner LUOYANG PULIKE WANTAI BIOTECH
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