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International Journal of Infectious Diseases: Volume 1, Number 4
Two-Step Polymerase Chain Reaction Assay for Detection of Yersinia Species in General and of Pathogenic Yersinia enterocolitica Strains Specifically
Paul T. Odinot, MSc; Jacques F.G.M. Meis, PhD, MD; Jo H.A.J. Curfs, PhD; Jacomina A.A. Hoogkamp-Korstanje, MD; and Willem J.G. Melchers, PhD

Int J Infect Dis 1997; 1(4):206-211.

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 Y. enterocolitica was specifically amplified in a nested PCR to identify pathogenic Y. enterocolitica.

Results: With this two-step PCR assay it was possible to detect Yersinieae in general as well as pathogenic Y. 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 Y. enterocolitica, and its ability to detect Y. enterocolitica in non-seeded samples.

Key Words: PCR detection, rats, 16S rRNA, virF, Yersinieae, Yersinia enterocolitica

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