Development of a recombinant OppA-based indirect hemagglutination test for the detection of antibodies against Haemophilus parasuis

文献类型： 外文期刊

第一作者： Chen, Shengli

作者： Chen, Shengli;Chu, Yuefeng;Zhao, Ping;He, Ying;Jian, Yingna;Liu, Yongsheng;Lu, Zhongxin

作者机构：

关键词： Haemophilus parasuis;OppA;Indirect hemagglutination test;Antibody detection

期刊名称：ACTA TROPICA （ 影响因子：3.112； 五年影响因子：3.002 ）

ISSN： 0001-706X

年卷期： 2015 年 148 卷

页码：

收录情况： SCI

摘要： An indirect hemagglutination (IHA) test that could detect antibodies against Haemophilus parasuis was developed. The full-length cDNA sequence of the oligopeptide permease ABC transporter membrane protein (OppA) gene was cloned, and inserted into the prokaryotic expression vector pET-30a(+) to construct recombinant plasmid pET-30a-OppA. The recombinant OppA protein was expressed partly in soluble form in Escherichia coil BL21 (DE3) and then purified by Ni2+ column. Furthermore, the recombinant OppA protein was used as an antigen to develop an IHA assay for detecting antibodies against H. parasuis. Results showed that this IHA test could detect species-specific antibodies against H. parasuis. Compared with currently available ELISA, the IHA test had a sensitivity of 85.0% and a specificity of 95.0%. The overall agreement between these two methods was 90.0%. The developed IHA test was used to evaluate the seroprevalence of H. parasuis in Hubei Province, China. The H. parasuis seroprevalence rate ranged from 5.5% to 26.2% in 325 tested clinical serum samples that were collected from three different pig farms in Hubei Province, China. The IHA test developed in this study will greatly contribute to the epidemiological surveys and immunization surveillance of H. parasuis. (C) 2015 Elsevier B.V. All rights reserved.

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