Molecular assays for detection of human metapneumovirus

Mackay, Ian M., Jacob, Kevin C., Woolhouse, Daniel, Waller, Katharine, Syrmis, Melanie W., Whiley, David M., Siebert, David J., Nissen, Michael and Sloots, Theo P. (2003) Molecular assays for detection of human metapneumovirus. Journal of Clinical Microbiology, 41 1: 100-105. doi:10.1128/JCM.41.1.100-105.2003

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Author Mackay, Ian M.
Jacob, Kevin C.
Woolhouse, Daniel
Waller, Katharine
Syrmis, Melanie W.
Whiley, David M.
Siebert, David J.
Nissen, Michael
Sloots, Theo P.
Title Molecular assays for detection of human metapneumovirus
Journal name Journal of Clinical Microbiology   Check publisher's open access policy
ISSN 1098-660X
Publication date 2003-01-01
Sub-type Article (original research)
DOI 10.1128/JCM.41.1.100-105.2003
Open Access Status File (Publisher version)
Volume 41
Issue 1
Start page 100
End page 105
Total pages 6
Editor Andrw B. Onderdonk
Place of publication Washington, DC, United States
Publisher American Society for Microbiology
Collection year 2003
Language eng
Abstract 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.
Keyword Metapneumovirus
Pcr Assay
Q-Index Code C1
Q-Index Status Provisional Code
Institutional Status Unknown

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Created: Tue, 14 Aug 2007, 19:14:34 EST