title: Development of a multiplex primer extension assay for rapid detection of Salmonella isolates of diverse serotypes.
abstract: Food-borne salmonellosis is a major manifestation of gastrointestinal disease in humans across the globe. Accurate and rapid identification methods could positively impact the identification of isolates, enhance outbreak investigation, and aid infection control. The SNaPshot multiplex system is a primer extension-based method that enables multiplexing of single nucleotide polymorphisms (SNPs). Here the method has been developed for the identification of five Salmonella serotypes, commonly detected in the United Kingdom, based on serotype-specific SNPs identified in the multilocus sequence typing (MLST) database of Salmonella enterica. The SNPs, in genes hemD, thrA, purE, and sucA, acted as surrogate markers for S. enterica serovars Typhimurium, Enteritidis, Virchow, Infantis, and Braenderup. The multiplex primer extension assay (MPEA) was conducted in two separate panels and evaluated using 152 Salmonella enterica isolates that were characterized by MLST. The MPEA was shown to be 100% specific and sensitive, within this collection of isolates. The MPEA is a sensitive and specific method for the identification and detection of Salmonella serotypes based upon SNPs seen in MLST data. The method can be applied in less than 6 h and has the potential to improve patient care and source tracing. The utility of the assay for identification of Salmonella serotypes directly from clinical specimens and food samples warrants further investigation.
authors.affiliation: Department of Medical Microbiology, School of Medicine, University of Manchester, Clinical Sciences Building, Manchester Royal Infirmary, Oxford Road, Manchester M13 9WL, United Kingdom.
authors.forename: Elloulu
authors.initials: E
authors.lastname: Ben-Darif
authors.forename: Francine
authors.initials: F
authors.lastname: Jury
authors.forename: Elizabeth
authors.initials: E
authors.lastname: De Pinna
authors.forename: E John
authors.initials: EJ
authors.lastname: Threlfall
authors.forename: Frederick J
authors.initials: FJ
authors.lastname: Bolton
authors.forename: Andrew J
authors.initials: AJ
authors.lastname: Fox
authors.forename: Mathew
authors.initials: M
authors.lastname: Upton
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D015373:Bacterial Typing Techniques; D004269:DNA, Bacterial / Q000235:genetics; D005517:Foodborne Diseases / Q000382:microbiology*; D005838:Genotype; D006801:Humans; D020641:Polymorphism, Single Nucleotide; D012480:Salmonella Infections / Q000175:diagnosis* / Q000382:microbiology; D019779:Salmonella enterica / Q000145:classification* / Q000235:genetics* / Q000302:isolation & purification; D012680:Sensitivity and Specificity; D012703:Serotyping; D006113:United Kingdom
chemical_list: D001426:Bacterial Proteins; D004269:DNA, Bacterial