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

doi:10.3390/v1010042

This article is

freely available
re-usable

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

Glenys R. Chidlow ^1,2^,* , Gerry B. Harnett ¹ , Geoffrey R. Shellam ² and David W. Smith ^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; in revised form: 2 June 2009 / Accepted: 5 June 2009 / Published: 8 June 2009

(This article belongs to the Special Issue Newly Identified Respiratory Viruses)

Download PDF Full-Text [169 KB, uploaded 8 June 2009 12:19 CET]

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

Article Statistics

Click here to load and display the download statistics.

Cite This Article

MDPI and ACS Style

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.

AMA Style

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(1):42-56.

Chicago/Turabian Style

Chidlow, Glenys R.; Harnett, Gerry B.; Shellam, Geoffrey R.; Smith, David W. 2009. "An Economical Tandem Multiplex Real-Time PCR Technique for the Detection of a Comprehensive Range of Respiratory Pathogens." Viruses 1, no. 1: 42-56.

Viruses EISSN 1999-4915 Published by MDPI Publishing, Basel, Switzerland RSS E-Mail Table of Contents Alert