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Viruses 2009, 1(1), 42-56; doi:10.3390/v1010042
Article

An Economical Tandem Multiplex Real-Time PCR Technique for the Detection of a Comprehensive Range of Respiratory Pathogens

1,2,* , 1
,
2
 and
1,2
1 Department of Microbiology, Pathwest Laboratory Medicine WA, QEII Medical Centre, Nedlands, WA 6009 Australia 2 School of Biomedical, Biomolecular and Chemical Sciences M502, University of Western Australia, Nedlands, WA 6009 Australia
* Author to whom correspondence should be addressed.
Received: 5 May 2009 / Revised: 2 June 2009 / Accepted: 5 June 2009 / Published: 8 June 2009
(This article belongs to the Special Issue Newly Identified Respiratory Viruses)
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Abstract

This study used real-time PCR assays to screen small sample volumes for a comprehensive range of 35 respiratory pathogens. Initial thermocycling was limited to 20 cycles to avoid competition for reagents, followed by a secondary real-time multiplex PCR. Supplementary semi-nested human metapneumovirus and picornavirus PCR assays were required to complete the acute respiratory pathogen profile. Potential pathogens were detected in 85 (70%) of pernasal aspirates collected from 121 children with acute respiratory symptoms. Multiple pathogens were detected in 29 (24%) of those samples. The tandem multiplex real-time PCR was an efficient method for the rapid detection of multiple pathogens.
Keywords: respiratory pathogens; tandem multiplex real-time PCR; mixed infections respiratory pathogens; tandem multiplex real-time PCR; mixed infections
This is an open access article distributed under the Creative Commons Attribution License (CC BY 3.0).
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Chidlow, G.R.; Harnett, G.B.; Shellam, G.R.; Smith, D.W. An Economical Tandem Multiplex Real-Time PCR Technique for the Detection of a Comprehensive Range of Respiratory Pathogens. Viruses 2009, 1, 42-56.

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