Preparation method of diagnostic kit for pig hepatitis E virus ELISA

A diagnostic kit and porcine hepatitis E technology, applied in the field of immunology, to achieve strong specificity, qualified stability, and good repeatability

Inactive Publication Date: 2010-06-09
SHANGHAI ACAD OF AGRI SCI
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  • Abstract
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AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to provide a preparation method of porci

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  • Preparation method of diagnostic kit for pig hepatitis E virus ELISA
  • Preparation method of diagnostic kit for pig hepatitis E virus ELISA
  • Preparation method of diagnostic kit for pig hepatitis E virus ELISA

Examples

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Embodiment 1

[0021] 1. Computer screening of porcine hepatitis E virus epitopes and synthesis of peptides

[0022] The computer software is used to screen the recognition epitopes from the corresponding proteins of the open reading frames ORF1, ORF2 and ORF3 of the whole genome of porcine hepatitis E virus. Epitope recognition is mainly based on the hydrophilicity, surface degree, flexibility of protein or polypeptide amino acid sequence, Chou-Fasman's helix, sheet and turn, Robson-Garnier's secondary structure and C- and N-terminus to predict Antigen comprehensive data (Antigenic Index, AI). Using existing software, such as: GCG (Madison, Wisconsin, USA) and DNAStar (Madison, Wisconsin, USA) to calculate AI and auxiliary recognition antigen regions from ORF1, ORF2 and ORF3 proteins of pig HEV. According to the amino acid sequences obtained from the known proteins of ORF1, ORF2 and ORF3 of porcine HEV, computer programs were used to estimate their antigenic regions (AI) (generally 10 to 1...

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Abstract

The invention discloses a preparation method of a diagnostic kit for pig hepatitis E virus ELISA, which comprises the following steps: (1) selecting antigen epitopes by a computer; (2) preparing antigens; (3) collecting serum; (4) establishing an indirect ELISA method; (5) verifying the sensibility and the specificity of the indirect ELISA method; (6) verifying the stability of the kit; and (7) carrying out clinical detection on a slaughterhouse. The method has the advantages of simplicity, high speed, high sensitivity, strong specificity, qualified stability and good repeatability. The kit prepared by the invention can effectively detect the hepatitis E virus of infected pigs, is suitable for detection of a large amount of pathogenetic samples, and can be used for quickly diagnosing the pig hepatitis E, preventing and controlling the spread of the pig hepatitis E virus and preventing the hepatitis E virus from spreading to people from pigs.

Description

technical field [0001] The invention belongs to the field of immunology, and in particular relates to a preparation method of a porcine hepatitis E virus ELISA diagnostic kit. Background technique [0002] Hepatitis E virus (Hepatitis E virus, HEV, referred to as hepatitis E) is a non-A-non-B hepatitis virus transmitted by fecal and oral routes. The virus can cause outbreaks in the population, mainly affecting young adults, and can cause a 20% fatality rate for pregnant women. From 1986 to 1988, Hepatitis E was prevalent in southern Xinjiang due to drinking water pollution, with a total of 119,280 cases and 707 deaths. It was the largest Hepatitis E epidemic in the world so far. In recent years, the incidence of hepatitis E has been increasing year by year. According to the statistics of the Ministry of Health of my country, the incidence of hepatitis E in 2003 increased by 40.4% compared with that in 2002. Therefore, the prevention and control of hepatitis E has been incl...

Claims

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Application Information

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IPC IPC(8): G01N33/576G01N33/531
Inventor 赵凯刘启文李震于瑞嵩唐雪明朱宏谭芙蓉吴潇王金斌陶世如蒋玲曦
Owner SHANGHAI ACAD OF AGRI SCI
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