Evaluation of recombinant outer membrane protein C based indirect enzyme-linked immunoassay for the detection of Salmonella antibodies in poultry

Abstract

Aim:To evaluate the efficacy of recombinant outer membrane proteinC (rOmpC) based enzyme-linked immunoassay (ELISA) for the diagnosis of salmonellosis in poultry.Materials and Methods:Three antigens were prepared, and the indirect ELISA was standardized using the antigens and the antiserum raised in chicken against Omp and rOmpC. Sera were collected from a total of 255 apparently healthy field chickens and screened for the presence of Salmonella antibodies by this ELISA.Results:The sodium dodecyl sulfate-polyacrylamide gel electrophoresis analysis of Omp revealed major polypeptides at 36, 42 and 52 kDa, and the rOmpC was evident by a single protein band of 43 kDa. The Omp and rOmpC antigen revealed an optimum concentration of 78 and 156 ng, respectively, in the assay, while the whole cell antigen gave an optimum reaction at a concentration of 106 organisms/ml. The test was found to be specific as it did not react with any of the antisera of seven other organisms. The developed ELISA detected Salmonella antibodies from 22 (8.62%) samples with rOmpC antigen, while 24 (9.41%) samples gave a positive reaction with both Omp and whole cell antigens.Conclusion:We suggest rOmpC based indirect ELISA as a suitable screening tool for serological monitoring of poultry flocks.

Characterization of Salmonella Enterica Isolated from Poultry Hatcheries and Commercial Broiler Chickens

“…Salmonella is a major foodborne pathogen responsible for many infectious diseases in animals and humans worldwide (Raguenaud et al, 2012: Manoj et al, 2015. In this study, a total of 8 Salmonella serotypes were isolated from Egyptian commercial broilers, dead chickens, and duck embryos.…”

Cloning, Sequencing and In Silico Characterization of OmpF Protein of Salmonella typhimurium for its Immune-Potential

“…Outer membrane proteins are good surface immunogens (Nandakumar et al, 1993), and are used in several studies to investigate the capability as a vaccine candidate in poultry (Meenakshi et al, 1999;Khan et al, 2003;Okamura et al, 2012;Prejit et al, 2013). Studies have also targeted in developments of diagnostic antigen with OMPs (Manoj et al, 2015). OmpC, OmpF, OmpD are the major outer membrane proteins of Salmonella (Singh et al, 2017).…”

A monoclonal antibody targeting the dimer interface of epidermal growth factor receptor (EGFR)

“…The purity and molecular weight of the monoclonal antibodies were determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The titer and affinity was calculated using an indirect ELISA [22,23] with OVA-EGFR 237-267 as the coating antigen.…”