Validation of a multiplex reverse transcriptase PCR ELISA for the detection of 19 respiratory tract pathogens

W Puppe, J Weigl, B Grondahl, M Knuf, S Rockahr, P von Bismarck, GI (Georgina) Aron, HGM Niesters, Ab Osterhaus, HJ Schmitt

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Introduction 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 (m-RT-PCR ELISA) to detect 19 different respiratory pathogens was developed and validated. Methods A total of 430 respiratory specimens were retrospectively tested in parallel by both the advanced 19-valent m-RT-PCR ELISA as well as by culture or individual RT-PCR assays used in clinical routine. Results The mean (median) sensitivity of the m-RT-PCR-ELISA in the retrospective test was 93.3% (95.1%; range 83.3-100 %), and the mean (median) specificity was 99.8 and 100 % (range 98.6-100 %), respectively. The mean positive predictive value was 99.3 % (range 934-100 %) and the mean negative predictive value was 95.3 % (range 98.4-100 %). Feasibility and clinical value of the 19-valent method was prospectively shown on 16,231 incoming clinical specimens from patients between 0 and 16 years of Conclusions The m-RT-PCR ELISA evaluated here improves the spectrum for diagnosing respiratory infections and is a feasible instrument for individual diagnostic and epidemiological studies.
Original languageUndefined/Unknown
Pages (from-to)77-91
Number of pages15
Issue number1
Publication statusPublished - 2013

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  • EMC MM-04-27-01

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