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J. Clin. Microbiol. doi:10.1128/JCM.02294-07
Copyright (c) 2008, American Society for Microbiology and/or the Listed Authors/Institutions. All Rights Reserved.

RespiFinder: a new multiparameter test to differentially identify 15 respiratory viruses

Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands; PathoFinder BV, Maastricht, The Netherlands; Department of Medical Microbiology and Infectious Diseases, Erasmus Medical Center Rotterdam, Rotterdam, The Netherlands

^* To whom correspondence should be addressed. Email: martin.reijans{at}pathofinder.com.

	Abstract

Broad-spectrum pathogen analysis in respiratory tract infections is becoming more relevant as the number of potential infectious agents is still increasing. Here we present the new multiparameter RespiFinder assay, which is based on the Multiplex Ligation-dependent Probe Amplification (MLPA) technology. This assay detects 15 respiratory viruses in one reaction. The MLPA reaction is preceded by a pre-amplification step which ensures the detection of both RNA and DNA viruses with the same specificity and sensitivity as individual monoplex real-time RT-PCRs. The RespiFinder was validated on 144 clinical samples and compared to cell culture and an RSV specific immuno-chromatography-assay (ICA). The RespiFinder showed a specificity and sensitivity of 98.2% and 100% for adenovirus, 96.4% and 100% for human metapneumovirus (hMPV), 98.2% and 100% for influenza A virus (InfA), 99.1% and 100% for parainfluenza virus 1 (PIV-1), 99.1% and 80% for PIV-3, 90.1% and 100% for rhinovirus, and 94.6% and 100% for respiratory syncytial virus (RSV), as compared to the cell culture results. Compared to the RSV-ICA the RespiFinder gave a specificity and sensitivity of 82.4% and 80%, respectively. PIV-2, PIV-4, InfB, InfA/H5N1 and coronavirus 229E were not detected in the clinical specimens tested. The use of the RespiFinder showed an increase of the diagnostic yield from 35.5% up to 60% compared to cell culture.

In conclusion, the RespiFinder assay provides a user-friendly and high-throughput tool for the simultaneous detection of 15 respiratory viruses with excellent overall performance statistics.