Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain
| Autor: | Qigai He, Huanchun Chen, Du Yujiao, Xuwang Cai, Yi Qi, Xiaojuan Xu, Yunbao Liu, Zhang Qinxue, Xiangru Wang, Yang Tian, Yuping Song |
|---|---|
| Jazyk: | angličtina |
| Rok vydání: | 2019 |
| Předmět: |
Serotype
Haemophilus Infections Swine Type V Secretion Systems Enzyme-Linked Immunosorbent Assay Microbiology Serology 03 medical and health sciences Haemophilus parasuis Antigen Bacterial Proteins Haemophilus Animals Pathogen 030304 developmental biology Antiserum Swine Diseases 0303 health sciences lcsh:Veterinary medicine General Veterinary biology 030306 microbiology General Medicine Gene Expression Regulation Bacterial biology.organism_classification RAPD Antibody detection Autotransporter Bacterial Vaccines biology.protein lcsh:SF600-1100 ELISA Antibody Research Article |
| Zdroj: | BMC Veterinary Research, Vol 15, Iss 1, Pp 1-9 (2019) BMC Veterinary Research |
| ISSN: | 1746-6148 |
| Popis: | Background Haemophilus parasuis is a commensal pathogen in the swine upper respiratory tract and causes Glässer’s disease. Surveillance, screening for infection, and vaccination response of H. parasuis is hindered by the lack of a rapid antibody detection method. Results In the present study, a monomeric autotransporter was identified as a novel antigen for developing an indirect ELISA. The autotransporter passenger domain (Apd) was expressed, purified, and demonstrated to be specific in ELISA and western blotting. Mouse antiserum of recombinant Apd (rApd) recognized native Apd in the 15 serotype reference strains and five non-typeable isolate stains, but showed no reaction with seven other bacterial pathogens. The rApd ELISA was optimized and validated using 67 serum samples with known background, including 27 positive sera from experimentally infected and vaccinated pigs along with 40 negative sera that had been screened with H. parasuis whole cell ELISA from clinically healthy herds. The rApd ELISA provided positive and negative percent agreements of 96.4 and 94.9%, respectively, and an AUC value of 0.961, indicating that the assay produced accurate results. Conclusion Apd was a universal antigen component among 15 serotype and non-typeable strains of H. parasuis and was also specific to this pathogen. The rApd ELISA could detect antibodies elicited by H. parasuis infection and vaccination, thereby exhibiting the potential to be applied for Glässer’s disease diagnosis, H. parasuis vaccination evaluation, and large-scale serological surveillance. |
| Databáze: | OpenAIRE |
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