This Article Full Text Full Text (PDF) Other Versions of this Article: JCM.02294-07v1 46/4/1232    most recent Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Reprints and Permissions Copyright Information Books from ASM Press MicrobeWorld Google Scholar Articles by Reijans, M. Articles by Simons, G. PubMed PubMed Citation Articles by Reijans, M. Articles by Simons, G.

RespiFinder: a New Multiparameter Test To Differentially Identify Fifteen Respiratory Viruses

Martin Reijans,^1* Gijs Dingemans,^1, Corné H. Klaassen,¹ Jacques F. Meis,¹ Judith Keijdener,² Brit Mulders,² Kimberly Eadie,³ Willem van Leeuwen,³ Alex van Belkum,³ Alphons M. Horrevorts,¹ and Guus Simons²

Department of Medical Microbiology and Infectious Diseases, Canisius Wilhelmina Hospital, Nijmegen, The Netherlands,¹ PathoFinder BV, Maastricht, The Netherlands,² Department of Medical Microbiology and Infectious Diseases, Erasmus Medical Center Rotterdam, Rotterdam, The Netherlands³

Received 29 November 2007/ Returned for modification 2 January 2008/ Accepted 25 January 2008

Broad-spectrum analysis for pathogens in patients with respiratory tract infections is becoming more relevant as the number of potential infectious agents is still increasing. Here we describe 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 preamplification step which ensures the detection of both RNA and DNA viruses with the same specificity and sensitivity as individual monoplex real-time reverse transcription-PCRs. The RespiFinder assay was validated with 144 clinical samples, and the results of the assay were compared to those of cell culture and a respiratory syncytial virus (RSV)-specific immunochromatography assay (ICA). Compared to the cell culture results, the RespiFinder assay showed specificities and sensitivities of 98.2% and 100%, respectively, for adenovirus; 96.4% and 100%, respectively, for human metapneumovirus; 98.2% and 100%, respectively, for influenza A virus (InfA); 99.1% and 100%, respectively, for parainfluenza virus type 1 (PIV-1); 99.1% and 80%, respectively, for PIV-3; 90.1% and 100%, respectively, for rhinovirus; and 94.6% and 100%, respectively, for RSV. Compared to the results of the RSV-specific ICA, the RespiFinder assay gave a specificity and a sensitivity of 82.4% and 80%, respectively. PIV-2, PIV-4, influenza B virus, InfA H5N1, and coronavirus 229E were not detected in the clinical specimens tested. The use of the RespiFinder assay resulted in an increase in the diagnostic yield compared to that obtained by cell culture (diagnostic yields, 60% and 35.5%, respectively). 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.

Published ahead of print on 6 February 2008.

Present address: PathoFinder B.V., Maastricht, The Netherlands.