Wo-Step Polymerase Chain Reaction Assay for Detection of Yersinia Species in General and of Pathogenic Yersinia enterocolitica Strains Specifically

Objectives: Detection of Yersinia enterocolitica in clinical samples is still not sensitive and fast enough. Polymerase chain reaction (PCR) offers the advantages of sensitivity, specificity, and rapidity. A two-step PCR assay to detect pathogenic Y: enterocolitica in infected tissue has been developed. Method: In the first step of the PCR assay, a general primer PCR amplification of the small subunit rDNA gene was used for the detection of a broad range of Yersinia species. In the second step, the virulence plasmid encoded virf-gene of \/: enterocolitica was specifically amplified in a nested PCR to identify pathogenic \/: enterocolitica. Results: With this two-step PCR assay it was possible to detect Yersinieae in general as well as pathogenic % enterocolitica specifically. The assay was then applied to detect Y: enterocolitica in spleens of experimentally infected rats. Discussion: This study shows the potential use of the PCR for the detection of pathogenic \/: enterocolitica, and its ability to detect Y: enterocolitica in non-seeded samples.