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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 which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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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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