title: Rapid identification of Salmonella enterica serovars, Typhimurium, Choleraesuis, Infantis, Hadar, Enteritidis, Dublin and Gallinarum, by multiplex PCR.
abstract: Salmonella enterica subsp. enterica poses a threat to both human and animal health, with more than 2500 serovars having been reported to date. Salmonella serovars are identified by slide and tube agglutination tests using O and H antigen-specific anti-sera, although this procedure is both labor intensive and time consuming. Establishment of a method for rapid screening of the major Salmonella serovars is therefore required. We have established multiplex polymerase chain reaction (m-PCR) assays for identification of seven serovars of Salmonella, i.e., Typhimurium, Choleraesuis, Infantis, Hadar, Enteritidis, Dublin and Gallinarum. Three serovar-specific genomic regions (SSGRs) of each serovar were selected using an approach in comparative genomics. The Salmonella-specific invA gene was used to confirm the genetic background of the organisms. The isolates tested were identified as a target serovar when the three selected SSGRs and invA were all positive for amplification. The specificity of each m-PCR assay was investigated using 118 serovars of Salmonella and 12 species of non-Salmonella strains. Although a small number of false-positive results were observed in the m-PCR assays used to identify Typhimurium, Choleraesuis, Enteritidis and Dublin for closely related serovars, false-negative results were not observed in any assays. These assays had sufficient specificity to identify the seven Salmonella serovars, and therefore, have the potential for use as rapid screening methods.
authors.affiliation: Safety Research Team, National Institute of Animal Health, 3-1-5 Kannondai, Tsukuba 305-0856, Japan. akiba@affrc.go.jp
authors.forename: Masato
authors.initials: M
authors.lastname: Akiba
authors.forename: Masahiro
authors.initials: M
authors.lastname: Kusumoto
authors.forename: Taketoshi
authors.initials: T
authors.lastname: Iwata
mesh_terms: D000818:Animals; D001431:Bacteriological Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D003951:Diagnostic Errors / Q000706:statistics & numerical data; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods*; D012480:Salmonella Infections / Q000175:diagnosis* / Q000382:microbiology; D012481:Salmonella Infections, Animal / Q000175:diagnosis* / Q000382:microbiology; D019779:Salmonella enterica / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial