title: Application of PCR-based serogrouping of selected Salmonella serotypes in Malaysia.
abstract: BACKGROUND
  Differentiation of Salmonella enterica into its serogroups is important for epidemiological study. The objective of the study was to apply a multiplex PCR targeting serogroups A, B, C1, D, E and Vi-positive strains of Salmonella enterica commonly found in Malaysia. A separate H-typing multiplex PCR which identified flagellar antigen "a", "b" or "d" was also optimized to confirm clinical serotypes, S. Paratyphi A and S. Typhi.
  
  
  METHODS
  Sixty-seven laboratory Salmonella enterica strains were tested. Six sets of primers targeting defined regions of the O antigen synthesis genes (rfb gene cluster) and Vi antigen gene (viaB) were selected and combined into a multiplex PCR for O-grouping. Four primers (H-for, Ha-rev, Hb-rev and Hd-rev) were used in the second step multiplex PCR for H-typing. The optimized mPCR assays were further evaluated with 58 blind-coded Salmonella strains.
  
  
  RESULTS
  The multiplex PCR results obtained showed 100% concordance to the conventionally typed serogroups. Validation with 58 blind coded Salmonella strains yield 100% accuracy and specificity.
  
  
  CONCLUSIONS
  Based on this study, PCR serogrouping proved to be a rapid, alternative method for further differentiation of Salmonella enterica.
authors.affiliation: Biomedical Science and Molecular Typing Laboratory, Institute of Biological Sciences, Faculty of Science, University of Malaya, Kuala Lumpur, Malaysia.
authors.forename: Bee Kim
authors.initials: BK
authors.lastname: Lim
authors.forename: Kwai Lin
authors.initials: KL
authors.lastname: Thong
mesh_terms: D015373:Bacterial Typing Techniques / Q000379:methods*; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000235:genetics; D005407:Flagella / Q000235:genetics; D006801:Humans; D008296:Malaysia; D019081:O Antigens / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods*; D011135:Polysaccharides, Bacterial / Q000235:genetics; D019779:Salmonella enterica / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification*; D012680:Sensitivity and Specificity; D012703:Serotyping
chemical_list: D017931:DNA Primers; D004269:DNA, Bacterial; D019081:O Antigens; D011135:Polysaccharides, Bacterial