title: An economical tandem multiplex real-time PCR technique for the detection of a comprehensive range of respiratory pathogens.
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.
authors: Department of Microbiology, Pathwest Laboratory Medicine WA, QEII Medical Centre, Nedlands, WA 6009 Australia.|School of Biomedical, Biomolecular and Chemical Sciences M502, University of Western Australia, Nedlands, WA 6009 Australia.
authors: Glenys R
authors: GR
authors: Chidlow
authors: Department of Microbiology, Pathwest Laboratory Medicine WA, QEII Medical Centre, Nedlands, WA 6009 Australia.
authors: Gerry B
authors: GB
authors: Harnett
authors: School of Biomedical, Biomolecular and Chemical Sciences M502, University of Western Australia, Nedlands, WA 6009 Australia.
authors: Geoffrey R
authors: GR
authors: Shellam
authors: Department of Microbiology, Pathwest Laboratory Medicine WA, QEII Medical Centre, Nedlands, WA 6009 Australia.|School of Biomedical, Biomolecular and Chemical Sciences M502, University of Western Australia, Nedlands, WA 6009 Australia.
authors: David W
authors: DW
authors: Smith