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

Abstract

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 coli BL21 (DE3) and then purified by Ni(2+) 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.

Keywords: Antibody detection; Haemophilus parasuis; Indirect hemagglutination test; OppA.