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Journal of Clinical Microbiology, January 2003, p. 100-105, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.100-105.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.

Molecular Assays for Detection of Human Metapneumovirus

Ian M. Mackay,1,2,3 Kevin C. Jacob,1 Daniel Woolhouse,1 Katharine Waller,1 Melanie W. Syrmis,1 David M. Whiley,1 David J. Siebert,4 Michael Nissen,2,4 and Theo P. Sloots1,2,3,4*

Clinical Virology Research Unit, Sir Albert Sakzewski Virus Research Centre, Royal Children's Hospital, Brisbane,1 Department of Paediatrics and Child Health,2 Clinical Medical Virology Centre, University of Queensland,3 Division of Microbiology, Queensland Health Pathology Service, Royal Brisbane Hospital Campus, Queensland, Australia4

Received 15 April 2002/ Returned for modification 19 August 2002/ Accepted 2 November 2002

The recent description of the respiratory pathogen human metapneumovirus (hMPV) has highlighted a deficiency in current diagnostic techniques for viral agents associated with acute lower respiratory tract infections. We describe two novel approaches to the detection of viral RNA by use of reverse transcriptase PCR (RT-PCR). The PCR products were identified after capture onto a solid-phase medium by hybridization with a sequence-specific, biotinylated oligonucleotide probe. The assay was applied to the screening of 329 nasopharyngeal aspirates sampled from patients suffering from respiratory tract disease. These samples were negative for other common microbial causes of respiratory tract disease. We were able to detect hMPV sequences in 32 (9.7%) samples collected from Australian patients during 2001. To further reduce result turnaround times we designed a fluorogenic TaqMan oligoprobe and combined it with the existing primers for use on the LightCycler platform. The real-time RT-PCR proved to be highly reproducible and detected hMPV in an additional 6 out of 62 samples (9.6%) tested during the comparison of the two diagnostic approaches. We found the real-time RT-PCR to be the test of choice for future investigation of samples for hMPV due to its speed, reproducibility, specificity, and sensitivity.

* Corresponding author. Mailing address: CVRU, SASVRC, Royal Children's Hospital, Herston Road, Herston, Queensland 4029, Australia. Phone: 61 3636 8833. Fax: 61 3636 1401. E-mail: t.sloots{at}mailbox.uq.edu.au.

Journal of Clinical Microbiology, January 2003, p. 100-105, Vol. 41, No. 1
0095-1137/03/$08.00+0     DOI: 10.1128/JCM.41.1.100-105.2003
Copyright © 2003, American Society for Microbiology. All Rights Reserved.



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