猪传染性胃肠炎病毒N蛋白原核表达和间接ELISA检测方法的建立

doi:10.16431/j.cnki.1671-7236.2016.09.012

Abstract

Abstract:

In the study,the recombinant expression plasmid pET28a-TGEV-N was constructed,and expression and purification of the TGEV N protein was completed.Through SDS-PAGE and Western blotting analysis,the N protein could be identified by transmissible gastroenteritis virus (TGEV) positive serum.Using the TGEV N protein as diagnosis antigen,we established an indirect ELISA method for detecting TGEV serum antibodies.The coefficients of variation of intro-batch and inter-batch duplicability test were less than 5%.The specificity was 100%,and the rate of false positive and the false negative rate were 0 and 5%,respectively.No cross reactions with seven porcine diseases positive serum were detected.Using this method,the clinical serum samples were detected.The results showed 100% coincidence rate compared with neutralization test.The method could detect TGEV antibody quickly and effectively,and had good repeatability and specificity,which laid the foundation for the development of standardized diagnostic kits.

Key words: transmissible gastroenteritis virus (TGEV); N protein; prokaryotic expression; indirect ELISA

CLC Number:

S852.65⁺1

GUO Hong-wei, ZHENG Ming, QIAO Hong-xing, MA Hui, ZHAO Xu-yong, WANG Yong-fen. Prokaryotic Expression and Development of an Indirect ELISA Detection Method of Porcine Transmissible Gastroenteritis Virus N Protein[J]. , 2016, 43(9): 2296-2301.

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Prokaryotic Expression and Development of an Indirect ELISA Detection Method of Porcine Transmissible Gastroenteritis Virus N Protein

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