title: 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.
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China.
authors.forename: Shengli
authors.initials: S
authors.lastname: Chen
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China.
authors.forename: Yuefeng
authors.initials: Y
authors.lastname: Chu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China.
authors.forename: Ping
authors.initials: P
authors.lastname: Zhao
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China.
authors.forename: Ying
authors.initials: Y
authors.lastname: He
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China.
authors.forename: Yingna
authors.initials: Y
authors.lastname: Jian
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China.
authors.forename: Yongsheng
authors.initials: Y
authors.lastname: Liu
authors.affiliation: State Key Laboratory of Veterinary Etiological Biology, Lanzhou Veterinary Research Institute, Chinese Academy of Agricultural Sciences, Xujiaping 1, Lanzhou 730046, Gansu, People's Republic of China. Electronic address: luzhongxin@caas.cn.
authors.forename: Zhongxin
authors.initials: Z
authors.lastname: Lu
mesh_terms: D000818:Animals; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000235:genetics; D002681:China / Q000453:epidemiology; D004797:Enzyme-Linked Immunosorbent Assay / Q000662:veterinary; D006192:Haemophilus Infections / Q000453:epidemiology / Q000382:microbiology / Q000662:veterinary*; D044137:Haemophilus parasuis / Q000235:genetics / Q000276:immunology / Q000302:isolation & purification*; D006386:Hemagglutination Tests / Q000662:veterinary; D026901:Membrane Transport Proteins / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000662:veterinary; D012680:Sensitivity and Specificity; D016036:Seroepidemiologic Studies; D013552:Swine; D013553:Swine Diseases / Q000453:epidemiology* / Q000382:microbiology
chemical_list: D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D026901:Membrane Transport Proteins; C037230:oligopeptide permease, Bacteria
keywords: Antibody detection; Haemophilus parasuis; Indirect hemagglutination test; OppA