Selective amplification of abequose and paratose synthase genes (rfb) by polymerase chain reaction for identification of Salmonella major serogroups (A, B, C2, and D)

Abstract

Many parts of the Salmonella rfb gene clusters which are responsible for biosynthesis of the oligosaccharide-repeating units of the O-antigenic lipopolysaccharide have recently been cloned and sequenced. On the basis of this knowledge, three sets of nucleotide primers were selected to target defined regions of the abequose and paratose synthase genes: rfbJ of Salmonella serogroup B, rfbJ of Salmonella serogroup C2, and rfbS of Salmonella serogroup D (also present in serogroup A). For good differentiation among these major serogroups, the primers were designed not only to give precise specificity in priming but also to give DNA products with different sizes in polymerase chain reactions (product sizes, ~720 bp for both serogroups A and D, ~820 bp for serogroup C2, and ~882 bp for serogroup B). In a polymerase chain reaction assay utilizing these rfb- specific primers, all of the 40 salmonellae belonging to serogroups B, C2, and D plus A were accurately identified among a total of 123 clinical isolates tested (including 55 salmonellae from 36 different serotypes and 68 strains from 10 other members of the family Enterobacteriaceae). No false- positive reactions were detected. The selected rfb gene sequences were proved for the first time to be useful DNA-based markers for identification of and differentiation among Salmonella serogroups A, B, C2, and D.