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

作者：Chidlow Glenys R^*; Harnett Gerry B; Shellam Geoffrey R; Smith David W

来源：Viruses-Basel, 2009, 1(1): 42-56.

DOI：10.3390/v1010042

• respiratory pathogens
• tandem multiplex real-time PCR
• mixed infections

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.

• 出版日期2009-6