Hepatitis E virus IgG antibody detection kit and preparation method and application thereof

An antibody detection and kit technology, applied in the field of hepatitis E virus IgG antibody kits, can solve the problems of high environmental cleanliness requirements and high dust, and achieve the effects of controlling the cost of reagents, improving sensitivity and improving specificity

Inactive Publication Date: 2016-03-30
SUZHOU SYM BIO LIFESCI CO LTD
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Although the detection sensitivity of time-resolved immunofluorescence analysis is greatly improved compared with traditional RIA (RadioImmunoassay, radioimmunoassay) or ELISA (Enzyme-linkedImmunoabsorbentassay, enzyme-linked immunoassay), compared with CLIA (ChemiluminescenceImmunoassay, chemiluminescent immunoassay), there are also There are certain advantages, but there are also some disadvantages, such as higher requirements on the cleanliness of the environment, because the dust in the air may cause the background to be high

Method used

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  • Hepatitis E virus IgG antibody detection kit and preparation method and application thereof
  • Hepatitis E virus IgG antibody detection kit and preparation method and application thereof
  • Hepatitis E virus IgG antibody detection kit and preparation method and application thereof

Examples

Experimental program
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Effect test

Embodiment 1

[0072] The preparation of embodiment 1 HEV-gene recombinant antigen coated plate

[0073] Source of HEV-gene recombinant antigen: purchased from Shanghai Biji Biotechnology Co., Ltd.

[0074] Proceed as follows:

[0075] (1) Print the barcode on the coated microporous reaction plate frame, and draw a red marking line on the raised part on the right side of the plate;

[0076] (2) After dissolving the HEV gene recombinant antigen in the coating buffer, soak the solid phase material and let it stand for 20 to 30 hours; wherein, the coating buffer is 0.05-0.1MpH8.0 (±0.2) PBS, and the coating The volume is 100 μL / well buffer solution, wherein, the buffer solution contains: 0.005-0.05% SDS; the coating working solution is added HEV gene recombinant antigen (150-400ng / mL) and 0.05%-0.5% (volume ratio) Antigen protection agent (β-mercaptoethanol) coating buffer; then add 100 μl / well into the microwell, let it stand at room temperature for 20-30 hours, and pay attention to prevent ...

Embodiment 2

[0081] The preparation of embodiment 2 europium markers

[0082] Source of mouse anti-human IgG monoclonal antibody: purchased from Beijing Xieshun Biotechnology Development Center

[0083] (1) Adjust the mouse anti-human IgG monoclonal antibody to 2.5mg / mL with ultrapure water, and dialyze overnight with a certain volume (1-5L) of dialysis solution 1 at a temperature of 20-25°C; It is 0.1mol / L, pH8.1±0.2 sodium bicarbonate buffer.

[0084] The dialysis time is 24h±2h, and the liquid is changed twice with an interval of more than 3 hours; after that, the dialysis solution 2 is used in the temperature range of 20-25°C; the dialysis buffer 2 is 0.1mol / L, pH 9.4± 0.2 CBS buffer. Dialyze for 24h±2h, change the medium twice, the time interval is more than 3 hours, and change the medium after get off work, and then dialyze overnight.

[0085] (2) At room temperature, according to the standard europium ratio of 1:3 (europium: protein), Eu-DTTA was completely dissolved in purified ...

Embodiment 3

[0087] Embodiment 3 detection application of hepatitis E IgG antibody

[0088] (1) Preparation of reagents

[0089]Mix 40mL concentrated lotion [Tris-HCl-NaCl (pH8.3±0.2) solution containing 0.2-1.0% Tween 20, diluted 40 times before use] and 960mL deionized water or distilled water in a clean lotion bottle , as a working cleaning solution.

[0090] Balance the negative and positive controls, sample diluent, analysis buffer, required number of microwell reaction strips and samples to be tested in the kit to 18-28°C), shake and mix the liquid reagents.

[0091] Prepare europium marker working solution: For each 12-well plate, take 75 μL europium marker (20×), add 1.5 mL analysis buffer [pH7.8±0.2 casein buffer, containing 0.878% NaCl, 0.5% Casein sodium salt, 0.606% Tris, 0.5% hydrochloric acid, 0.00074% EDTA-Na 2 , 0.2% TritonX-100, 3% goat serum, 0.005% cockscomb and 0.05% NaN 3 】, mix thoroughly (1:20 dilution). Prepare within 30 minutes before the second shaking reacti...

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Abstract

The invention discloses a hepatitis E virus IgG antibody detection kit. The kit includes the following components: a solid material coated with a hepatitis E virus (HEV) gene recombinant antigen, a mouse anti human IgG monoclonal antibody labeled with a signal generation substance, a concentrated washing solution, an enhancing liquid, an analysis buffer liquid, a sample diluent, a positive control and a negative control. In addition, the invention also provides a preparation method of the kit. The preparation method includes the following steps: a first step, preparing the solid material coated with the HEV gene recombinant antigen; and a second step, labeling a mouse anti human IgG monoclonal antibody with the signal generation substance. In addition, the invention also discloses an application of the kit in detection of a hepatitis E virus IgG antibody. The detection kit overcomes the defect of labeling antibodies with macromolecules (such as enzymes), has the advantages of high sensitivity, good stability, low cost and the like, can significantly improve the specificity, sensitivity and stability of detection of the hepatitis E virus IgG antibody, and besides, significantly reduces the cost.

Description

technical field [0001] The invention relates to a hepatitis E virus (HEV) IgG antibody kit and a detection method thereof, in particular to a method for qualitatively detecting hepatitis E virus IgG antibody in a human serum sample based on the non-competitive immunoassay principle of indirect labeling and its corresponding detection kits. Background technique [0002] Hepatitis E is an acute gastrointestinal infectious disease caused by Hepatitis E Virus (HEV) and transmitted through fecal and oral routes. It is mainly prevalent in developing countries and regions such as Asia, Africa and Latin America. Hepatitis E and Hepatitis A are very common in my country and other developing countries. Ten years ago, outbreaks of Hepatitis A and Hepatitis E occurred in Shanghai and Xinjiang respectively, with more than 100,000 cases. The two are very similar in terms of clinical symptoms, transmission route and outcome, but the incidence of hepatitis E is mainly in young and middle-a...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): G01N33/577
Inventor 彭会军
Owner SUZHOU SYM BIO LIFESCI CO LTD
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