Novel real-time multiplex PCR for rapid identification of pathogenic Yersinia species
(2017) ECCMID 2017 — Location: Vienne (22.April.2017)
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- Abstract
- Background: Among the 18 species of the genus Yersinia, only three species are responsible for human diseases: Y. pestis is the causative agent of plague and has become uncommon in Europe. Y. pseudotuberculosis and Y. enterocolitica biotype 1B, 2, 3, 4, 5 are responsible for gastrointestinal diseases and are found occasionally from stool cultures in bacteriology laboratories. The virulence of these bacteria is related to the presence of virulence factors located in the chromosome and in virulence plasmids. The National Reference Center (NRC) for Yersinia currently performs identification of Yersinia strains received from clinical specimen, with the aim to determine the pathogenicity based on the species, biotype and serotype determination. Due to the complexity of these gold-standard techniques, time to mean time-to-result is 4 days. In our experience, only 45% of Yersinia species isolated from clinical samples are pathogenic. Material/methods: We designed a real-time multiplex PCR allowing rapid identification of pathogenic Yersinia species. The technique targets three genes. hrpA is a pan-Yersinia gene and is used as positive control. inV is a gene specific for Y. pseudotuberculosis and Y. pestis. yopM is present in the virulence plasmid of all pathogenic Yersinia species. This combination of three targets allows to rapidly distinguish non-pathogenic species from those responsible for human disease (figure 1). The multiplex PCR was performed on purified DNA using the Lightcycler© 96 system (Roche Diagnostics). Results: We performed the multiplex PCR in parallel with Gold Standard techniques on a panel of Yersinia species, 13 enteric species and 19 consecutive strains received by the NRC for identification (Table 1). Only two discrepancies were observed, as the PCR did not detect the hrpA gene of the nonpathogenic Y. massiliensis and Y. frederiksenii strains. These discrepancies did not have a clinical impact. All PCR test results were available on the same day. Conclusions: We developed a new multiplex PCR method allowing the rapid identification of pathogenic Yersinia species. This should allow to accelerate the response for clinicians awaiting the clinical relevance of a positive Yersinia culture.
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André, E., De Sany, P., Darricades, M., Goeminne, L., Michiels, T., Janssens, M., Wauters, G., & Delmée, M. (2017). Novel real-time multiplex PCR for rapid identification of pathogenic Yersinia species. ECCMID 2017, Vienne. https://hdl.handle.net/2078.5/49158