A molecular scheme for Yersinia enterocolitica patho-serotyping derived from genome-wide analysis.
Yersinia enterocolitica is a food-borne, gastro-intestinal pathogen with world-wide distribution. Only 11 serotypes have been isolated from patients, with O:3, O:9, O:8 and O:5,27 being the serotypes most commonly associated with human yersiniosis. Serotype is an important characteristic of Y. enterocolitica strains, allowing differentiation for epidemiology, diagnosis and phylogeny studies. Conventional serotyping, performed by slide agglutination, is a tedious and laborious procedure whose interpretation tends to be subjective, leading to poor reproducibility. Here we present a PCR-based typing scheme for molecular identification and patho-serotyping of Y. enterocolitica. Genome-wide comparison of Y. enterocolitica sequences allowed analysis of the O-antigen gene clusters of different serotypes, uncovering their formerly unknown genomic locations, and selection of targets for serotype-specific amplification. Two multiplex PCRs and one additional PCR were designed and tested on various reference strains and isolates from different origins. Our genotypic assay proved to be highly specific for identification of Y. enterocolitica species, discrimination between virulent and non-virulent strains, distinguishing the main human-related serotypes, and typing of conventionally untypeable strains. This genotyping scheme could be applied in microbiology laboratories as an alternative or complementary method to the traditional phenotypic assays, providing data for epidemiological studies.
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology, Ludwig Maximilians-University, Munich, Germany.
Debora
D
Garzetti
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology, Ludwig Maximilians-University, Munich, Germany.
Rosa
R
Susen
Robert Koch Institute, Wernigerode Branch, Division Enteropathogenic Bacteria and Legionella, National Reference Centre for Salmonellae and Other Bacterial Enteric Pathogens, Wernigerode, Germany.
Angelika
A
Fruth
Robert Koch Institute, Wernigerode Branch, Division Enteropathogenic Bacteria and Legionella, National Reference Centre for Salmonellae and Other Bacterial Enteric Pathogens, Wernigerode, Germany.
Erhard
E
Tietze
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology, Ludwig Maximilians-University, Munich, Germany.
Jürgen
J
Heesemann
Max von Pettenkofer-Institute for Hygiene and Medical Microbiology, Ludwig Maximilians-University, Munich, Germany. Electronic address: rakin@mvp.uni-muenchen.de.
Alexander
A
Rakin
D016680:Genome, Bacterial; D005838:Genotype; D060005:Genotyping Techniques; D006801:Humans; D058889:Molecular Typing / Q000379:methods*; D005810:Multigene Family; D019081:O Antigens / Q000235:genetics; D016133:Polymerase Chain Reaction / Q000379:methods; D012703:Serotyping; D015009:Yersinia Infections / Q000175:diagnosis / Q000382:microbiology; D015008:Yersinia enterocolitica / Q000145:classification* / Q000235:genetics*
D019081:O Antigens
Genome-wide comparison; Molecular patho-serotyping; O-antigen; Yersinia enterocolitica