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EB virus VCA-IgA antibody detection reagent and preparation method thereof

An Epstein-Barr virus and antibody detection technology, applied in measurement devices, disease diagnosis, instruments, etc., can solve the problems of competition interference, reduction of sensitivity and specificity of detection results, etc., to improve sensitivity and specificity, wide application range, and reduce operation effect of error

Active Publication Date: 2015-01-14
中山生物工程有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0009] The above two colloidal gold method detection reagents all use EB virus antigen-coated detection line, and the antigen of the detection line can react with all antibodies such as EB virus IgG, IgA, IgM in the test sample, because IgG is the main component of immunoglobulin in serum , accounting for about 75% of the total immunoglobulin content in serum, which is 4 to 7 times that of the target IgA antibody, which can seriously compete and interfere with the combination of the target IgA and the coated EB virus antigen, resulting in a decrease in the sensitivity and specificity of the test results

Method used

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  • EB virus VCA-IgA antibody detection reagent and preparation method thereof
  • EB virus VCA-IgA antibody detection reagent and preparation method thereof
  • EB virus VCA-IgA antibody detection reagent and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Embodiment 1 detection reagent preparation condition screening

[0039] 1. Selection of colloidal gold particles

[0040] 1.1 Principle: Colloidal gold is prepared according to the redox reaction between chloroauric acid and trisodium citrate under boiling conditions. The particle size of the colloidal gold can be changed and controlled by adjusting the addition ratio of chloroauric acid and trisodium citrate.

[0041] 1.2 Preparation method

[0042] Add 1ml of 1% chloroauric acid solution to 100ml of double-distilled water and boil, add different amounts of 1% trisodium citrate solution under stirring conditions, continue to boil for 5 minutes to prepare colloidal gold particles of different sizes, and cool naturally for later use. Colloidal gold of different sizes was used to label avidin, and the company’s internal quality control product was used as the research material for detection. The results are shown in Table 1 and Table 2.

[0043] Table 1. Selection ex...

Embodiment 2

[0092] Embodiment 2. Reagent preparation

[0093] 1. Raw material requirements

[0094] 1.1 Recombinant Epstein-Barr virus VCA antigen

[0095] Manufacturer: Hong Kong Shennong Co., Ltd.

[0096] Product Name: Recombinant Epstein-Barr Virus VCA Antigen, Recombinant Epstein-Barr Virus NA1 Antigen

[0097] Batch number: 20131224E

[0098] Appearance: colorless transparent liquid;

[0099] Concentration and purity requirements: the concentration is greater than 2.0 mg / ml, determined by SDS-PAGA, and only one band exists under the condition of 10 microliters of sample.

[0100] 1.2 Mouse anti-human IgA antibody

[0101] Manufacturer: Hangzhou Longji Biotechnology Co., Ltd.

[0102] Product name: mouse anti-human IgA antibody

[0103] Batch number: 20131105

[0104] Appearance: colorless transparent liquid;

[0105] Concentration and purity requirements: the concentration is greater than 2.0 mg / ml, and it is determined by SDS-PAGA. The sample volume is 10 microliters and...

Embodiment 3

[0160] Embodiment 3.EB virus VCA-IgA antibody detection kit (colloidal gold method) preparation

[0161]Epstein-Barr virus VCA IgA antibody detection kit (colloidal gold method), including mouse anti-human IgA monoclonal antibody coated on the NC membrane detection line, biotin coated on the quality control line, and biotin-labeled antibody coated on the antigen pad Colloidal gold-labeled avidin coated on the recombinant Epstein-Barr virus VCA antigen and the gold label pad. Mouse anti-human IgA monoclonal antibody is a colorless transparent liquid with a concentration greater than 2.0mg / ml. It is determined by SDS-PAGA. There are two bands under the condition of 10μl of sample loading, and the coating concentration is 1.4mg / ml; avidin is Colorless transparent liquid, the concentration is greater than 2.0mg / ml, determined by SDS-PAGA, there is only one band under the condition of 10μl sample volume, the labeling concentration is 25μg / ml; biotin is a colorless transparent liq...

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Abstract

The invention provides an EB virus VCA-IgA antibody detection reagent which at least comprises a reactive film, an antigen pad and a gold-labeled pad, wherein a detection line and a quality control line are marked on the reactive film; the detection line contains a mouse anti-human IgA monoclonal antibody; the quality control line contains biotin; the antigen pad contains a biotin-labeled recombinant EB virus VCA antigen; and the gold-labeled pad contains an avidin complex labeled by colloidal gold. The detection kit disclosed by the invention has the characteristics of rapidness, simplicity, accuracy and high accuracy.

Description

technical field [0001] The invention relates to the field of biotechnology, in particular to an antibody detection reagent. [0002] The invention also relates to a kit containing the detection reagent and a preparation method. Background technique [0003] Epstein-Barr virus (EBv), also known as human herpesvirus 4 (Human herpesvirus 4 (HHV-4)). In 1964, Epstein and Barr successfully established Burkitt African children's lymphoma cells through in vitro suspension culture for the first time in 1964, and observed herpes virus particles in the cell smears of the established strains with an electron microscope, thinking that the virus is a variety of malignant tumors (such as One of the causes of nasopharyngeal carcinoma), it mainly infects the epithelial cells and B lymphocytes of the human oropharynx. Most of the patients with nasopharyngeal carcinoma in southern China have detected the presence of Epstein-Barr virus genome. Epstein-Barr virus is widely distributed, mostl...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577G01N33/531G01N33/569
CPCG01N33/543G01N33/56994G01N2800/14G01N2800/7028
Inventor 唐安洲王胜岚叶为民张哲宋小冬李峰
Owner 中山生物工程有限公司
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