Development of a blocking ELISA for detection of antibodies against avian hepatitis E virus

文献类型： 外文期刊

第一作者： Liu, Baoyuan

作者： Liu, Baoyuan;Zhao, Qin;Sun, Yani;Wang, Xinjie;Zhao, Jinan;Du, Taofeng;Wang, Chengbao;Xiao, Shuqi;Mu, Yang;Zhang, Gaiping;Zhou, En-Min;Liu, Baoyuan;Zhao, Qin;Sun, Yani;Wang, Xinjie;Zhao, Jinan;Du, Taofeng;Wang, Chengbao;Xiao, Shuqi;Mu, Yang;Zhou, En-Min;Zhang, Gaiping;Luo, Jianxuri;Hsu, Walter H.

作者机构：

关键词： Avian hepatitis E virus;Blocking ELISA;Monoclonal antibody;ORF2 protein

期刊名称：JOURNAL OF VIROLOGICAL METHODS （ 影响因子：2.014； 五年影响因子：2.001 ）

ISSN：

年卷期：

页码：

收录情况： SCI

摘要： A blocking enzyme-linked immunosorbent assay (bELISA) was developed for the detection of immunoglobulin G antibodies against avian hepatitis E virus (REV). In the bELISA, the coating antigen was a truncated protein containing C-terminal 268-amino acid region of ORF2 from an avian HEV strain isolated in China (CaHEV) and blocking antibody was a monoclonal antibody (mAb) 1H5 recognizing the epitope within amino acids 384-414 in the C-terminal 268-amino acid region. The concentration of blocking mAb 1H5 was determined as that yielded an OD450nm value of 1.0 for binding to the coating antigen and the antigen concentration and serum dilution were optimized using a checkerboard titration. A cut-off value of 20.7% at the mean percent inhibition plus 3 standard deviations was determined by testing 265 negative sera. The bELISA had a sensitivity of 98.3% by testing 116 positive sera from chickens infected experimentally with CaHEV and had no cross-reaction with other anti-avian virus antibodies. The compliance rates of the bELISA with indirect ELISA and Western blot were 83.7% and 93.3%, respectively, by testing 300 field chicken sera. These results suggested that the bELISA developed in this study can be used for detection of antibodies against avian REV and showed high reproducibility compared with indirect ELISA and Western blot methods

分类号： R37

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