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EB (epstein-barr) virus VCA/NA1-IgA antibody joint detection reagent and preparation method thereof

A joint detection technology of Epstein-Barr virus, applied in the direction of measuring devices, disease diagnosis, instruments, etc., can solve the problems of reduced sensitivity and specificity of detection results, competition interference, etc. Effect

Active Publication Date: 2015-01-21
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

[0012] 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 (epstein-barr) virus VCA/NA1-IgA antibody joint detection reagent and preparation method thereof
  • EB (epstein-barr) virus VCA/NA1-IgA antibody joint detection reagent and preparation method thereof
  • EB (epstein-barr) virus VCA/NA1-IgA antibody joint detection reagent and preparation method thereof

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Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1 detection reagent preparation condition screening

[0042] 1. Selection of colloidal gold particles

[0043] 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.

[0044] 1.2 Preparation method

[0045] 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.

[0046] Table 1. Selection ex...

Embodiment 2

[0099] Embodiment 2. Reagent preparation

[0100] 1. Raw material requirements

[0101] 1.1 Recombinant Epstein-Barr virus VCA antigen and NA1 antigen

[0102] Manufacturer: Hong Kong Shennong Co., Ltd.

[0103] Product Name: Recombinant Epstein-Barr Virus VCA Antigen, Recombinant Epstein-Barr Virus NA1 Antigen

[0104] Batch number: 20131224E(VCA), 20140126E(NA1)

[0105] Appearance: colorless transparent liquid;

[0106] 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.

[0107] 1.2 Mouse anti-human IgA antibody

[0108] Manufacturer: Hangzhou Longji Biotechnology Co., Ltd.

[0109] Product name: mouse anti-human IgA antibody

[0110] Batch number: 20131105

[0111] Appearance: colorless transparent liquid;

[0112] Concentration and purity requirements: the concentration is greater than 2.0 mg / ml, and it is determined by SDS-PAGA. Th...

Embodiment 3

[0175] Embodiment 3.EB virus VCA / NA1-IgA antibody combined detection kit (colloidal gold method) preparation

[0176] (1) Preparation of streaked NC membrane: Dilute mouse anti-human IgA monoclonal antibody to a coating concentration of 1.4 mg / mL with phosphate buffer, dilute biotin to a coating concentration of 2.2 mg / mL, and use The membrane machine draws the two coating solutions onto the NC membrane respectively, and saves the coated NC membrane after drying;

[0177] (2) Preparation of avidin gold-labeled pad: Colloidal gold-labeled biotin solution was prepared by coupling avidin with a labeling concentration of 30 μg / mL to colloidal gold, centrifuged at 10,000 r / min for 30 min, and discarded the supernatant , add the colloidal gold conjugate dilution to 80% of the original volume, and then use the mixed solution to press 60μl / m 2 Soak and smear to make a gold standard pad, and store the gold standard pad dry;

[0178] (3) Preparation of the antigen pad: according to ...

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Abstract

The invention provides an EB (epstein-barr) virus VCA / NA1-IgA antibody joint detection reagent. The EB virus VCA / NA1-IgA antibody joint detection reagent at least comprises a reaction membrane, an antigen pad and a gold label pad, wherein a detection line and a quality control line are labeled on the reaction membrane; the detection line contains a rat anti-human IgA monoclonal antibody; the quality control line contains biotin; the antigen pad contains a recombinant EB virus VCA antigen labeled with the biotin and a recombinant EB virus NA1 antigen labeled with the biotin; and the gold label pad contains an avidin compound containing a colloid gold label. The joint detection reagent provided by the invention is used for carrying out joint detection on VCA and NA1 so that the cost can be saved; the application and the popularization of the detection reagent are facilitated and the detection cost is reduced; and the EB virus VCA / NA1-IgA antibody joint detection reagent has the characteristics of rapidness, simplicity and convenience, accuracy and high sensitivity.

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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IPC IPC(8): G01N33/577
CPCG01N33/54306G01N33/56994G01N2800/14G01N2800/7028
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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