Herbert Tomaso, 11Bundeswehr Institute of Microbiology, Munich, Germany
Daniela Jacob, 22Robert Koch-Institut, Zentrum für Biologische Sicherheit, Hochpathogene mikrobielle Erreger, Berlin, Germany
Meike Eickhoff, 33QIAGEN Hamburg GmbH, Hamburg, Germany
Holger C. Scholz, 44Bundeswehr Institute of Microbiology, Munich, Germany
Sascha Al Dahouk, 55Department of Internal Medicine III, RWTH Aachen University, Aachen, Germany
Mireille M. Kattar, 66Molecular Infectious Diseases Diagnostics, Department of Pathology and Laboratory Medicine, American University of Beirut, Beirut, Lebanon
Udo Reischl, 77Institute of Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany
Helga Plicka, 88BMLV/RD-ARWT, ABCUT, Mödling, Austria
Jaran Strand Olsen, 99Norwegian Defense Research Establishment, Division for Protection, Kjeller, Norway
Simo Nikkari, 1010Center for Biothreat Preparedness, and BC-Defense and Environmental Health Unit, Center for Military Medicine, Helsinki, Finland
Pirjo Matero, 1111Center for Biothreat Preparedness, and BC-Defense and Environmental Health Unit, Center for Military Medicine, Helsinki, Finland
Christian Beuret, 1212Biology Spiez Laboratory, Federal Department of Defense, Civil Protection and Sports, Federal Office for Civil Protection, Spiez, Switzerland
Andrea Ciammaruconi, 1313Health Corps, Italian Army, Histology and Molecular Biology Section, Army Medical and Veterinary Research Center, Roma, Italy
Florigio Lista, 1414Health Corps, Italian Army, Histology and Molecular Biology Section, Army Medical and Veterinary Research Center, Roma, Italy
Jean-Luc Gala, 1515Center for Applied Molecular Technologies, Defense Laboratories Department, Belgian Armed Forces, Brussels, Belgium
Hermann Broll, 1616Bundesinstitut für Risikobewertung, Berlin, Germany
Bernd Appel, 1717Bundesinstitut für Risikobewertung, Berlin, Germany
Ricela E. Sellek Cano, 1818Unidad Biológica-NBQ, Fábrica Nacional “La Marañosa”, Spain
Maria del Carmen Ybarra de Villavicencio, 1919Unidad Biológica-NBQ, Fábrica Nacional “La Marañosa”, Spain
Martien Broekhuijsen, 2020TNO Defense, Security and Safety, Rijswijk, The Netherlands
Alexander Indra, 2121AGES – Institut für medizinische Mikrobiologie und Hygiene, Wien, Austria
Roger Petersen, 2222TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany
Heinrich Neubauer, 2323Friedrich Loeffler Institut, Jena, Germany
Corresponding author: Dr. Herbert Tomaso, Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, 80937 Munich, Germany Phone: +49-89-3168-3933, Fax: +49-89-3168-3292,
Citation Information. Clinical Chemistry and Laboratory Medicine. Volume 46, Issue 9, Pages 1239–1244, ISSN (Online) 1437-4331, ISSN (Print) 1434-6621, DOI: 10.1515/CCLM.2008.251, Available online: 21/07/2008, September 2008
Publication History: Received: 12/3/2008; accepted: 12/5/2008; published online: 21/07/2008
Abstract
Background: Yersinia pestis (Y. pestis) is a zoonotic bacterium mainly circulating among rodents and their fleas. Transmission to humans can cause bubonic, pneumonic or septicemic plague with a high case-fatality rate. Therefore, rapid and reliable diagnostic tools are crucial. The objective of this study was to assess the inter-laboratory reproducibility of in-house developed real-time PCR assays for the identification of Y. pestis.
Methods: A total of four samples of quantified Y. pestis DNA and two blank samples were sent blinded to 14 laboratories. To standardize the procedures, oligonucleotides were provided and the same instrument platform and a commercial mastermix were used. The participants were requested to report their results including cycle threshold and melting temperature values.
Results: All participating laboratories were able to perform the real-time PCR assays according to the protocols provided and identified the samples containing Y. pestis DNA correctly. Significant differences between the reference laboratory and participating laboratories were observed in cycle threshold values and melting temperatures. This, however, did not adversely affect the interpretation of results.
Conclusions: Our real-time PCR system proved to be highly reproducible and has the potential of complementing the diagnostic tools for rapid identification of Y. pestis isolates. Further steps of validation are needed to determine diagnostic accuracy and predictive values with clinical samples.
Clin Chem Lab Med 2008;46:1239–44.
Keywords Yersinia pestis, real-time PCR
