Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain
Abstract 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 stud...
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| Format: | Article |
| Language: | English |
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BMC
2019-12-01
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| Series: | BMC Veterinary Research |
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| Online Access: | https://doi.org/10.1186/s12917-019-2128-x |
| _version_ | 1819173870814363648 |
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| author | Yunbao Liu Yujiao Du Yuping Song Yang Tian Yi Qi Qinxue Zhang Qigai He Xiangru Wang Huanchun Chen Xuwang Cai Xiaojuan Xu |
| author_facet | Yunbao Liu Yujiao Du Yuping Song Yang Tian Yi Qi Qinxue Zhang Qigai He Xiangru Wang Huanchun Chen Xuwang Cai Xiaojuan Xu |
| author_sort | Yunbao Liu |
| collection | DOAJ |
| description | Abstract 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. |
| first_indexed | 2024-12-22T20:29:57Z |
| format | Article |
| id | doaj.art-66a62db2fe0441cbbae7269b5b03842d |
| institution | Directory Open Access Journal |
| issn | 1746-6148 |
| language | English |
| last_indexed | 2024-12-22T20:29:57Z |
| publishDate | 2019-12-01 |
| publisher | BMC |
| record_format | Article |
| series | BMC Veterinary Research |
| spelling | doaj.art-66a62db2fe0441cbbae7269b5b03842d2022-12-21T18:13:38ZengBMCBMC Veterinary Research1746-61482019-12-011511910.1186/s12917-019-2128-xDevelopment and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domainYunbao Liu0Yujiao Du1Yuping Song2Yang Tian3Yi Qi4Qinxue Zhang5Qigai He6Xiangru Wang7Huanchun Chen8Xuwang Cai9Xiaojuan Xu10State Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityThe cooperative Innovation Center for Sustainable Pig Production, Key Laboratory of Preventive Veterinary Medicine in Hubei ProvinceState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityState Key Laboratory of Agricultural Microbiology, College of Veterinary Medicine, Huazhong Agricultural UniversityAbstract 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.https://doi.org/10.1186/s12917-019-2128-xHaemophilus parasuisELISAAntibody detectionAutotransporter |
| spellingShingle | Yunbao Liu Yujiao Du Yuping Song Yang Tian Yi Qi Qinxue Zhang Qigai He Xiangru Wang Huanchun Chen Xuwang Cai Xiaojuan Xu Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain BMC Veterinary Research Haemophilus parasuis ELISA Antibody detection Autotransporter |
| title | Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain |
| title_full | Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain |
| title_fullStr | Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain |
| title_full_unstemmed | Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain |
| title_short | Development and application of an antibody detection ELISA for Haemophilus parasuis based on a monomeric autotransporter passenger domain |
| title_sort | development and application of an antibody detection elisa for haemophilus parasuis based on a monomeric autotransporter passenger domain |
| topic | Haemophilus parasuis ELISA Antibody detection Autotransporter |
| url | https://doi.org/10.1186/s12917-019-2128-x |
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