Preparation of a monoclonal antibody based sandwich enzyme-linked immunosorbent assay for Salmonella

A double antibody sandwich enzyme-linked immunosorbent assay (ELISA) was developed using monoclonal antibodies (mAbs) as a rapid, economical alternative to culture isolation procedures for detection of Salmonella. Through cell fusion technology, five mAbs were screened out, in which A2C6D3 as capture antibody and E3C5E8 as detection antibody were selected for the development of ELISA method. This assay detected all Salmonella serovars, which included representative serovars of serogroups B, C, D, and E and gave negative results for all non-Salmonella species tested. The ELISA had a specificity and sensitivity of 100% and 91%, respectively, and a kappa value of 0.93 relative to the culture methods. Such a sandwich ELISA has the potential to be used in the implementation of the pathogen reduction and hazard analysis critical control point systems as well as in clinical laboratories.