title: A rapid screen of broth enrichments for Salmonella enterica serovars enteritidis, Hadar, Heidelberg, and Typhimurium by Using an allelotyping multiplex PCR that targets O- and H-antigen alleles.
abstract: Salmonella continues to cause significant foodborne outbreaks, best illustrated with recent outbreaks associated with peanut butter, raw tomatoes, and serrano peppers. To ascertain the likely source of the outbreak, bacterial typing is essential to this process. While PCR has become an important detection tool for pathogens in foods, PCR can also identify strain differences by targeting gene(s) or sequences exhibiting polymorphisms or variability in its distribution within the bacterial population. Over 2,500 Salmonella enterica serovars identified based on antigenic differences in lipopolysaccharide and flagellin have been identified to date. We developed an allelotyping PCR scheme that identifies the O and H alleles associated with S. enterica serovars Enteritidis, Hadar, Heidelberg, Typhimurium, and others, with the same antigen alleles but in different O- and H-allele combinations (e.g., S. enterica Kentucky), and validated it as a screen to identify samples contaminated with these Salmonella serovars. We correctly identified poultry samples containing S. enterica serovars Enteritidis, Kentucky, and Typhimurium from our multiplex screen of primary enrichments of environmental drag swabs. PCR agreed well (kappa values = 0.81 to 1.0) with conventional serotyping methods used to type salmonellae isolated from primary enrichment. Coupled with Salmonella-specific PCR, such as invA, this allelotyping PCR could prove useful in the identification of Salmonella and specific S. enterica serovars present in foods or the environment and could decrease the time and cost associated with conventional serotyping methods.
authors.affiliation: Department of Population Health, College of Veterinary Medicine, The University of Georgia, Athens, Georgia 30602, USA.
authors.forename: Yang
authors.initials: Y
authors.lastname: Hong
authors.forename: Tongrui
authors.initials: T
authors.lastname: Liu
authors.forename: Margie D
authors.initials: MD
authors.lastname: Lee
authors.forename: Charles L
authors.initials: CL
authors.lastname: Hofacre
authors.forename: Marie
authors.initials: M
authors.lastname: Maier
authors.forename: David G
authors.initials: DG
authors.lastname: White
authors.forename: Sherry
authors.initials: S
authors.lastname: Ayers
authors.forename: Lihua
authors.initials: L
authors.lastname: Wang
authors.forename: Roy
authors.initials: R
authors.lastname: Berghaus
authors.forename: John
authors.initials: J
authors.lastname: Maurer
mesh_terms: D000483:Alleles; D000942:Antigens, Bacterial / Q000235:genetics*; D015373:Bacterial Typing Techniques; D005506:Food Contamination / Q000032:analysis; D019081:O Antigens / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D019779:Salmonella enterica / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D012477:Salmonella enteritidis / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D012486:Salmonella typhimurium / Q000145:classification / Q000235:genetics / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012703:Serotyping
chemical_list: D000942:Antigens, Bacterial; C022863:H antigen; D019081:O Antigens