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Background  Classical  Salmonella  serotyping is an expensive and time consuming process that requires implementing a battery of O and H antisera to detect 2,541 different  Salmonella enterica  serovars. For these reasons, we developed a rapid multiplex polymerase chain reaction (PCR)-based typing scheme to screen for the prevalent  S. enterica  serovars Enteritidis, Hadar, Heidelberg, and Typhimurium.    Results  By analyzing the nucleotide sequences of the genes for O-antigen biosynthesis including  wb a operon and the central variable regions of the H1 and H2 flagellin genes in  Salmonella , designated PCR primers for four multiplex PCR reactions were used to detect and differentiate  Salmonella  serogroups A/D1, B, C1, C2, or E1; H1 antigen types i, g, m, r or z 10 ; and H2 antigen complexes, I: 1,2; 1,5; 1,6; 1,7 or II: e,n,x; e,n,z 15 . Through the detection of these antigen gene allele combinations, we were able to distinguish among  S. enterica  serovars Enteritidis, Hadar, Heidelberg, and Typhimurium. The assays were useful in identifying  Salmonella  with O and H antigen gene alleles representing 43 distinct serovars. While the H2 multiplex could discriminate between unrelated H2 antigens, the PCR could not discern differences within the antigen complexes, 1,2; 1,5; 1,6; 1,7 or e,n,x; e,n,z 15 , requiring a final confirmatory PCR test in the final serovar reporting of  S. enterica .    Conclusion  Multiplex PCR assays for detecting specific O and H antigen gene alleles can be a rapid and cost-effective alternative approach to classical serotyping for presumptive identification of  S. enterica  serovars Enteritidis, Hadar, Heidelberg, and Typhimurium.

bmc microbiology

Rapid screening of Salmonella entericaserovars Enteritidis, Hadar, Heidelberg and Typhimurium using a serologically-correlative allelotyping PCR targeting the O and H antigen alleles