A proteomic approach to the development of DIVA ELISA distinguishing pigs infected with Salmonella Typhimurium and pigs vaccinated with a Salmonella Typhimurium-based inactivated vaccine

BMC Veterinary Research

Methodology Article

Marcel Kosina¹  Hana Kudlackova²  Martin Faldyna²  Radek Tesarik²  Jan Matiasovic²  Jan Gebauer³  Alena Osvaldova⁴  Kamil Kovarcik⁵ 

[1] Bioveta a. s., Komenskeho212/12, 683 23, Ivanovice na Hane, Czech Republic;Department of Immunology, Veterinary Research Institute, Hudcova296/70, 62100, Brno, Czech Republic;Department of Immunology, Veterinary Research Institute, Hudcova296/70, 62100, Brno, Czech Republic;Department of Experimental Biology, Faculty of Science, Masaryk University, Kotlarska267/2, 611 37, Brno, Czech Republic;Department of Immunology, Veterinary Research Institute, Hudcova296/70, 62100, Brno, Czech Republic;Faculty of Veterinary Medicine, University of Veterinary and Pharmaceutical Sciences, Palackeho 1/3, 612 42, Brno, Czech Republic;Department of Virology, Veterinary Research Institute, Hudcova296/70, 62100, Brno, Czech Republic;

关键词: Porcine;    Salmonella;    DIVA vaccine;    Recombinant protein;    Mass spectrometry;   

DOI  :  10.1186/s12917-016-0879-1

 received in 2016-04-21, accepted in 2016-11-03,  发布年份 2016

来源: Springer

【 摘 要 】

BackgroundSalmonella enterica serovar Typhimurium is one of the most common enteropathogenic bacteria found in pigs in Europe. In our previous work, we demonstrated the protective effects in suckling piglets when their dams had been vaccinated with an S. Typhimurium-based inactivated vaccine. This study is focused on a procedure leading to serological discrimination between vaccinated and infected pigs. As we supposed, distinct environment during natural infection and in bacterial cultures used for vaccine preparation led to a slightly different spectrum of expressed S. Typhimurium proteins. The examination of porcine antibodies produced after the experimental infection with S. Typhimurium or after vaccination with S. Typhimurium-based inactivated vaccine by affinity chromatography and mass spectrometry revealed differences in antibody response applicable for serological differentiation of infected from vaccinated animals.ResultsAntibodies against Salmonella SipB, SipD and SseB proteins were detected at much higher levels in post-infection sera in comparison with control and post-vaccination sera. On the other hand, proteins BamB, OppA and a fragment of FliC interacted with antibodies from post-vaccination sera with a much higher intensity than from control and post-infection sera. In addition, we constructed ELISA assays using post-infection antigen - SipB protein and post-vaccination antigen - FliC-fragment and evaluated them on a panel of individual porcine sera.ConclusionsThe analysis of antibody response of infected and vaccinated pigs by proteomic tools enabled to identify S. Typhimurium antigens useful for distinguishing infected from vaccinated animals. This approach can be utilized in other challenges where DIVA vaccine and a subsequent serological assay are required, especially when genetic modification of a vaccine strain is not desirable.

【 授权许可】

CC BY   
© The Author(s). 2016