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Preliminary validation of real-time PCR assays for the identification of Yersinia pestis

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dc.contributor.author Tomaso H.
dc.contributor.author Jacob D.
dc.contributor.author Eickhoff M.
dc.contributor.author Scholz H.C.
dc.contributor.author Al Dahouk S.
dc.contributor.author Kattar M.M.
dc.contributor.author Reischl U.
dc.contributor.author Plicka H.
dc.contributor.author Olsen J.S.
dc.contributor.author Nikkari S.
dc.contributor.author Matero P.
dc.contributor.author Beuret C.
dc.contributor.author Ciammaruconi A.
dc.contributor.author Lista F.
dc.contributor.author Gala J.-L.
dc.contributor.author Broll H.
dc.contributor.author Appel B.
dc.contributor.author Sellek Cano R.E.
dc.contributor.author Ybarra De Villavicencio M.D.C.
dc.contributor.author Bro
dc.contributor.editor
dc.date 2008
dc.date.accessioned 2017-10-05T15:59:21Z
dc.date.available 2017-10-05T15:59:21Z
dc.date.issued 2008
dc.identifier 10.1515/CCLM.2008.251
dc.identifier.isbn
dc.identifier.issn 14346621
dc.identifier.uri http://hdl.handle.net/10938/18960
dc.description.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. © 2008 by Walter de Gruyter.
dc.format.extent
dc.format.extent Pages: (1239-1244)
dc.language English
dc.publisher BERLIN
dc.relation.ispartof Publication Name: Clinical Chemistry and Laboratory Medicine; Publication Year: 2008; Volume: 46; no. 9; Pages: (1239-1244);
dc.relation.ispartofseries
dc.relation.uri
dc.source Scopus
dc.subject.other GENBANK: AE013964, AF053947, AL117189, JO2459, M27820, X92727
dc.title Preliminary validation of real-time PCR assays for the identification of Yersinia pestis
dc.type Article
dc.contributor.affiliation Tomaso, H., Bundeswehr Institute of Microbiology, Munich, Germany, Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, 80937 Munich, Germany
dc.contributor.affiliation Jacob, D., Robert Koch-Institut, Zentrum für Biologische Sicherheit, Hochpathogene Mikrobielle Erreger, Berlin, Germany
dc.contributor.affiliation Eickhoff, M., QIAGEN Hamburg GmbH, Hamburg, Germany
dc.contributor.affiliation Scholz, H.C., Bundeswehr Institute of Microbiology, Munich, Germany
dc.contributor.affiliation Al Dahouk, S., Department of Internal Medicine III, RWTH Aachen University, Aachen, Germany
dc.contributor.affiliation Kattar, M.M., Molecular Infectious Diseases Diagnostics, Department of Pathology and Laboratory Medicine, American University of Beirut, Beirut, Lebanon
dc.contributor.affiliation Reischl, U., Institute of Medical Microbiology and Hygiene, University of Regensburg, Regensburg, Germany
dc.contributor.affiliation Plicka, H., BMLV-RD-ARWT, ABCUT, Mödling, Austria
dc.contributor.affiliation Olsen, J.S., Norwegian Defense Research Establishment, Division for Protection, Kjeller, Norway
dc.contributor.affiliation Nikkari, S., Center for Biothreat Preparedness, BC-Defense and Environmental Health Unit, Center for Military Medicine, Helsinki, Finland
dc.contributor.affiliation Matero, P., Center for Biothreat Preparedness, BC-Defense and Environmental Health Unit, Center for Military Medicine, Helsinki, Finland
dc.contributor.affiliation Beuret, C., Biology Spiez Laboratory, Federal Department of Defense, Federal Office for Civil Protection, Spiez, Switzerland
dc.contributor.affiliation Ciammaruconi, A., Health Corps, Histology and Molecular Biology Section, Army Medical and Veterinary Research Center, Roma, Italy
dc.contributor.affiliation Lista, F., Health Corps, Histology and Molecular Biology Section, Army Medical and Veterinary Research Center, Roma, Italy
dc.contributor.affiliation Gala, J.-L., Center for Applied Molecular Technologies, Defense Laboratories Department, Belgian Armed Forces, Brussels, Belgium
dc.contributor.affiliation Broll, H., Bundesinstitut für Risikobewertung, Berlin, Germany
dc.contributor.affiliation Appel, B., Unidad Biológica-NBQ, Fábrica Nacional la Marañosa, Spain
dc.contributor.affiliation Sellek Cano, R.E., Unidad Biológica-NBQ, Fábrica Nacional la Marañosa, Spain
dc.contributor.affiliation Ybarra De Villavicencio, M.D.C., Unidad Biológica-NBQ, Fábrica Nacional la Marañosa, Spain
dc.contributor.affiliation Broekhuijsen, M., TNO Defense, Security and Safety, Rijswijk, Netherlands
dc.contributor.affiliation Indra, A., AGES - Institut für Medizinische Mikrobiologie und Hygiene, Wien, Austria
dc.contributor.affiliation Petersen, R., TIB MOLBIOL Syntheselabor GmbH, Berlin, Germany
dc.contributor.affiliation Neubauer, H., Friedrich Loeffler Institut, Jena, Germany
dc.contributor.authorAddress Tomaso, H.; Bundeswehr Institute of Microbiology, Neuherbergstrasse 11, 80937 Munich, Germany; email: herbert.tomaso@web.de
dc.contributor.authorCorporate University: American University of Beirut Medical Center; Faculty: Faculty of Medicine; Department: Pathology and Laboratory Medicine;
dc.contributor.authorDepartment Pathology and Laboratory Medicine
dc.contributor.authorDivision
dc.contributor.authorEmail herbert.tomaso@web.de
dc.contributor.authorFaculty Faculty of Medicine
dc.contributor.authorInitials Tomaso, H
dc.contributor.authorInitials Jacob, D
dc.contributor.authorInitials Eickhoff, M
dc.contributor.authorInitials Scholz, HC
dc.contributor.authorInitials Al Dahouk, S
dc.contributor.authorInitials Kattar, MM
dc.contributor.authorInitials Reischl, U
dc.contributor.authorInitials Plicka, H
dc.contributor.authorInitials Olsen, JS
dc.contributor.authorInitials Nikkari, S
dc.contributor.authorInitials Matero, P
dc.contributor.authorInitials Beuret, C
dc.contributor.authorInitials Ciammaruconi, A
dc.contributor.authorInitials Lista, F
dc.contributor.authorInitials Gala, JL
dc.contributor.authorInitials Broll, H
dc.contributor.authorInitials Appel, B
dc.contributor.authorInitials Cano, RES
dc.contributor.authorInitials de Villavicencio, MDY
dc.contributor.authorInitials Broekhuijsen, M
dc.contributor.authorInitials Indra,
dc.contributor.authorOrcidID
dc.contributor.authorReprintAddress Tomaso, H (reprint author), Bundeswehr Inst Microbiol, Neuherbergstr 11, D-80937 Munich, Germany.
dc.contributor.authorResearcherID
dc.contributor.authorUniversity American University of Beirut Medical Center
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dc.description.citedCount 9
dc.description.citedTotWOSCount 9
dc.description.citedWOSCount 8
dc.format.extentCount 6
dc.identifier.articleNo
dc.identifier.coden CCLMF
dc.identifier.pubmedID 18783342
dc.identifier.scopusID 51849136926
dc.identifier.url
dc.publisher.address GENTHINER STRASSE 13, D-10785 BERLIN, GERMANY
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dc.relation.ispartofConferenceCode
dc.relation.ispartofConferenceDate
dc.relation.ispartofConferenceHosting
dc.relation.ispartofConferenceLoc
dc.relation.ispartofConferenceSponsor
dc.relation.ispartofConferenceTitle
dc.relation.ispartofFundingAgency
dc.relation.ispartOfISOAbbr Clin. Chem. Lab. Med.
dc.relation.ispartOfIssue 9
dc.relation.ispartOfPart
dc.relation.ispartofPubTitle Clinical Chemistry and Laboratory Medicine
dc.relation.ispartofPubTitleAbbr Clin. Chem. Lab. Med.
dc.relation.ispartOfSpecialIssue
dc.relation.ispartOfSuppl
dc.relation.ispartOfVolume 46
dc.source.ID WOS:000259929300006
dc.type.publication Journal
dc.subject.otherAuthKeyword Real-time PCR
dc.subject.otherAuthKeyword Yersinia pestis
dc.subject.otherChemCAS
dc.subject.otherIndex article
dc.subject.otherIndex bacterium identification
dc.subject.otherIndex bacterium isolate
dc.subject.otherIndex controlled study
dc.subject.otherIndex diagnostic accuracy
dc.subject.otherIndex nonhuman
dc.subject.otherIndex nucleotide sequence
dc.subject.otherIndex priority journal
dc.subject.otherIndex real time polymerase chain reaction
dc.subject.otherIndex validation process
dc.subject.otherIndex Yersinia pestis
dc.subject.otherIndex Biological Warfare Agents
dc.subject.otherIndex Laboratories
dc.subject.otherIndex Polymerase Chain Reaction
dc.subject.otherIndex Reproducibility of Results
dc.subject.otherIndex Time Factors
dc.subject.otherIndex Yersinia pestis
dc.subject.otherIndex Bacteria (microorganisms)
dc.subject.otherIndex Rodentia
dc.subject.otherIndex Siphonaptera (fleas)
dc.subject.otherIndex Yersinia pestis
dc.subject.otherKeywordPlus CHAIN-REACTION ASSAY
dc.subject.otherKeywordPlus PLAGUE
dc.subject.otherKeywordPlus SEQUENCE
dc.subject.otherWOS Medical Laboratory Technology


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