Sensitive and rapid immunoassays for Salmonella enteritidis

In order to develop more sensitive, specific, and rapid immunoassays to detect Salmonella enteritidis in food supplies, we have applied various approaches by using several different antibody preparations. Utilizing ELISA in both a plate and immunodot assay, we employed (i) a polycolonal rabbit antiserum to a boiled suspension of the organism; (ii) a monoclonal antibody to the cell surface of the bacterium; and (iii) mouse antisera to two oligosaccharides each containing the rare sugar tyvelose, and exhibited by S. enteritidis, a member of the group D salmonellae. We showed that the polyclonal antiserum and monoclonal antibody IG-10 to the cell surface could specifically detect from 10(2) to 10(3) organisms in a 10-microl sample in the plate and immunodot assay. Both assays are read in 4-5 hr. Further, in the mice immunized to the trisaccharide, (alpha-D-galactose-alpha-tyvelose-alpha-D-mannose), as well as those mice immunized to the tetrasaccharide, (alpha-D-galactose-alpha-tyvelose-alpha-D-mannose-alpha-L-rhamnose), specificity to tyvelose was determined by inhibition studies. The inhibitors of the antisera to the trisaccharide included the single sugar tyvelose conjugated to bovine serum albumin (BSA), a tetrasaccharide in which tyvelose is excluded, but contains alpha-D-galactose, alpha-ascarose, alpha-D-mannose, and alpha-L-rhamnose (conjugated to BSA), and others. The inhibition studies suggest that the mouse antisera are specific for tyvelose and also contain antibodies for mannose and rhamnose. The antibodies that have been made to the unique sugar tyvelose should improve the specificity in assays for S. enteritidis.