title: [Development of molecular PCR-RFLP test for identification of the epidemic strain of Y. enterocolitica bioserotype 1B/O8 circulating in Poland since 2004].
abstract: BACKGROUND
  Highly pathogenic Y. enterocolitica bioserotype 1B/O8 is considered to be an important etiological agent of yersiniosis in Poland. Infections caused by Y. enterocolitica 1B/O8 became an important public health problem in Poland, especially because of their high potential of virulence and the unknown source of the bacteria. Y. enterocolitica 1B/O8 isolates recovered in Poland are genetically highly related and constitute single epidemic sensu stricto strain. The aim of the present study was to develop a time- and money-effective molecular assay for rapid identification of pathogenic Y. enterocolitica 1B/O8 isolates belonging to the epidemic strain.
  
  
  METHODS
  In the first stage we performed a multiplex-PCR for four genetic markers: ail, ystA, irp1 and 16S rDNA sequence. In the next stage we designed a duplex-PCR-RFLP assay with BtsI endonuclease to detect/identify specific variant of an ysrR gene that is characteristic for epidemic strain of Y. enterocolitica 1B/O8 strain. The assay was tested against a panel of a consisted of a variety Yersinia enterocolitica and Y. pseudotuberculosis strains.
  
  
  RESULTS
  All the tested Y. enterocolitica 1B/O8 strains were positive for all the genetic markers in multiplex-PCR assay what distinguished them from other tested Yersinia strains. In duplex-PCR-RFLP test all tested isolates of the epidemic strain were negative for ysrR digestion with BtsI endonuclease, while all tested reference strains of Y. enterocolitica 1B/O8 were positive.
  
  
  CONCLUSIONS
  The assay developed in this study was two-stage/two-step molecular test efficiently distinguishing wild-type and the epidemic Y. enterocolitica 1B/O8 strain. Such test can be a useful screening tool for clinical, veterinary and food diagnostics, as well as for the purposes of epidemiological investigation.
authors.forename: Tomasz
authors.initials: T
authors.lastname: Wołkowicz
authors.forename: Natalia
authors.initials: N
authors.lastname: Wolaniuk
authors.forename: Katarzyna
authors.initials: K
authors.lastname: Zacharczuk
authors.forename: Rafał
authors.initials: R
authors.lastname: Gierczyński
authors.forename: Natalia
authors.initials: N
authors.lastname: Rokosz
authors.forename: Waldemar
authors.initials: W
authors.lastname: Rastawicki
mesh_terms: D004196:Disease Outbreaks; D005819:Genetic Markers / Q000235:genetics; D006801:Humans; D011044:Poland / Q000453:epidemiology; D016133:Polymerase Chain Reaction / Q000379:methods*; D012150:Polymorphism, Restriction Fragment Length; D012703:Serotyping; D013045:Species Specificity; D014774:Virulence / Q000235:genetics; D015009:Yersinia Infections / Q000453:epidemiology / Q000382:microbiology*; D015008:Yersinia enterocolitica / Q000145:classification* / Q000235:genetics / Q000302:isolation & purification* / Q000472:pathogenicity
chemical_list: D005819:Genetic Markers