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. 2003 Mar;69(3):1810-6.
doi: 10.1128/AEM.69.3.1810-1816.2003.

PCR detection of virulence genes in Yersinia enterocolitica and Yersinia pseudotuberculosis and investigation of virulence gene distribution

P Thoerner  1 , C I Bin Kingombe, K Bögli-Stuber, B Bissig-Choisat, T M Wassenaar, J Frey, T Jemmi
Affiliations

PCR detection of virulence genes in Yersinia enterocolitica and Yersinia pseudotuberculosis and investigation of virulence gene distribution

P Thoerner et al. Appl Environ Microbiol. 2003 Mar.

Abstract

PCR-based assays were developed for the detection of plasmid- and chromosome-borne virulence genes in Yersinia enterocolitica and Yersinia pseudotuberculosis, to investigate the distribution of these genes in isolates from various sources. The results of PCR genotyping, based on 5 virulence-associated genes of 140 strains of Y. enterocolitica, were compared to phenotypic tests, such as biotyping and serotyping, and to virulence plasmid-associated properties such as calcium-dependent growth at 37 degrees C and Congo red uptake. The specificity of the PCR results was validated by hybridization. Genotyping data correlated well with biotype data, and most biotypes resulted in (nearly) homogeneous genotypes for the chromosomal virulence genes (ystA, ystB, and ail); however, plasmid-borne genes (yadA and virF) were detected with variable efficiency, due to heterogeneity within the bacterial population for the presence of the virulence plasmid. Of the virulence genes, only ystB was present in biotype 1A; however, within this biotype, pathogenic and apathogenic isolates could not be distinguished based on the detection of virulence genes. Forty Y. pseudotuberculosis isolates were tested by PCR for the presence of inv, yadA, and lcrF. All isolates were inv positive, and 88% of the isolates contained the virulence plasmid genes yadA and lcrF. In conclusion, this study shows that genotyping of Yersinia spp., based on both chromosome- and plasmid-borne virulence genes, is feasible and informative and can provide a rapid and reliable genotypic characterization of field isolates.

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Figures

FIG. 1.
FIG. 1.
Diagram of the distribution of Y. enterocolitica strains included in this study. (A) Strains sorted by biotype and source. (B) Strains stacked by genotype for each isolation source. Reference strains are not included.
FIG. 2.
FIG. 2.
Distribution of virulence genes and biotypes in serotypes O:3 and O:9 of Y. enterocolitica. (A) Distribution of the virulence genes in both serotypes. (B) Distribution of biotypes for the two serotypes
FIG. 3.
FIG. 3.
Diagram of the distribution of Y. enterocolitica genotypes, as determined by PCR, in different biotypes. (A) Distribution of genotypes in biotypes isolated from human sources. (B) Distribution of genotypes in biotypes isolated from nonhuman sources.
See this image and copyright information in PMC

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