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Title of Journal: Infection

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Abbravation: Infection

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Springer-Verlag

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DOI

10.1002/bip.360100606

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1439-0973

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Validation of a multiplex reverse transcriptase PC

Authors: W Puppe J Weigl B Gröndahl M Knuf S Rockahr P von Bismarck G Aron H G M Niesters A D M E Osterhaus HJ Schmitt
Publish Date: 2012/07/31
Volume: 41, Issue: 1, Pages: 77-91
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Abstract

Since acute respiratory tract infections inflict a high burden of disease in children worldwide a multiplex reverse transcription polymerase chain reaction combined with a microwell hybridization assay mRTPCR–ELISA to detect 19 different respiratory pathogens was developed and validatedThe mean median sensitivity of the mRTPCR–ELISA in the retrospective test was 933 951 range 833–100  and the mean median specificity was 998 and 100  range 986–100  respectively The mean positive predictive value was 993  range 934–100  and the mean negative predictive value was 953  range 984–100  Feasibility and clinical value of the 19valent method was prospectively shown on 16231 incoming clinical specimens from patients between 0 and 16 years of age with acute respiratory tract infections admitted to pediatric hospitals or private practices from October 2003 to June 2010 in three regions in Germany Kiel Mainz Freiburg Freiburg to June 2007 only At least one microorganism was detected in 10765 of 16231 663  clinical specimens 5044 RV 1999 RSV 1286 AV 944 EV 737 seasonal IVA 173 pandemic IVA H1N12009 899 MPV 518 CV 383 PIV3 268 PIV1 259 Mpn 205 IVB 164 PIV2 144 PIV4 103 Bp 29 Cpn and 29 Bpp while reovirus and Lpn were not present in these specimens from a pediatric population More than one organism could be detected in 134  of the specimensWe appreciate the excellent technical assistance of B Reckewitz N Esly and E BudoGuetaifi and we thank I Chaloupka Institute for Medical Microbiology Department of Virology TU Munich Germany E Jacobs and R Dumke Institute of Medical Microbiology University Hospital Dresden Germany RP Verkooyen Erasmus University Rotterdam the Netherlands M Maaß Institute for Medical Microbiology University of Lübeck Germany E Staube Institute of Clinical Microbiology University Hospital Jena Germany Dr B Schweiger RKI Berlin Germany for providing several specimens and strains used in this study C Drosten Institute of Virology University of Bonn Medical Centre Bonn Germany R Fouchier Erasmus Medical Center Rotterdam the Netherlands J Ziebuhr University of Giessen Germany and Dr Ijzerman Haarlem the Netherlands for providing and retesting of the Legionella pneumophila strains This work was supported by the Bundesministerium für Bildung und Forschung BMBF via the Deutsches Zentrum für Luft und Raumfahrt eV by the research grant 01KI9910/1 for “PIDARInet” a pediatric infectious diseases network on acute respiratory tract infections


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