Development of recombinant flagellar antigens for serological detection of Salmonella enterica serovars enteritidis, hadar, heidelberg, and typhimurium in poultry
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Accurate and fast serological detection of harmful Salmonella in poultry is a major concern of food safety. Common Salmonella serotypes responsible for human associated foodborne outbreaks are Enteritidis, Hadar, Heidelberg, and Typhimurium. Recombinant flagellin for phase 1 (H1) and phase 2 (H2) were cloned into Pinpoint-T® protein expression vector and purified to be used as antigens in an ELISA. An ELISA was developed for the serological detection of S. enterica Enteritidis in this study. 500 ng of purified recombinant g,m antigen and a 1:64 dilution was determined to be optimal for testing field serum samples through checkerboard titration analysis. A negative baseline cutoff was calculated to be an optical density (OD) of 0.35. This new ELISA eliminates common cross reactivity inherent in lipopolysaccharide (LPS) or whole cell antigens by focusing specifically on the H1 and H2 phase combination of flagellin found on common Salmonella serotypes present in poultry.