Detection of Yersinia pestis using real-time PCR in patients with suspected bubonic plague.

Julia M Riehm (1) , Lila Rahalison (2) , Holger C Scholz (1) , Bryan Thoma (1) , Martin Pfeffer (3) , Léa Mamiharisoa Razanakoto (2) , Sascha Al Dahouk (4, 5) , Heinrich Neubauer (6) , Herbert Tomaso (6)

1 Bundeswehr Institute of Microbiology
2 Laboratoire Central de la Peste (CNR)
3 Institute of Animal Hygiene and Veterinary Public Health
4 RWTH - Rheinisch-Westfälische Technische Hochschule Aachen University
5 BfR - Bundesinstitut für Risikobewertung - Federal Institute for Risk Assessment
6 IBIZ - Institute of Bacterial Infections and Zoonoses

Yersinia (Y.) pestis, the causative agent of plague, is endemic in natural foci of Asia, Africa, and America. Real-time PCR assays have been described as rapid diagnostic tools, but so far none has been validated for its clinical use. In a retrospective clinical study we evaluated three real-time PCR assays in two different assay formats, 5'-nuclease and hybridization probes assays. Lymph node aspirates from 149 patients from Madagascar with the clinical diagnosis of bubonic plague were investigated for the detection of Y. pestis DNA. Results of real-time PCR assays targeting the virulence plasmids pPCP1 (pla gene), and pMT1 (caf1, Ymt genes) were compared with an F1-antigen immunochromatographic test (ICT) and cultivation of the organism. Out of the 149 samples an infection with Y. pestis was confirmed by culture in 47 patients while ICT was positive in 88 including all culture proven cases. The best real-time PCR assay was the 5'-nuclease assay targeting pla which was positive in 120 cases. In conclusion, the 5'-nuclease assay targeting pla can be recommended as diagnostic tool for establishing a presumptive diagnosis when bubonic plague is clinically suspected.

Keywords: Real-time PCR; Yersinia pestis; Bubonic plague; F1-antigen; Madagascar

Domains: Life Sciences [q-bio]

Julia M Riehm, Lila Rahalison, Holger C Scholz, Bryan Thoma, Martin Pfeffer, et al.. Detection of Yersinia pestis using real-time PCR in patients with suspected bubonic plague.. Molecular and Cellular Probes, 2011, 25 (1), pp.8-12. ⟨10.1016/j.mcp.2010.09.002⟩. ⟨pasteur-00836493⟩