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.
Viruses. 2009 Jun;1(1):42-56. doi: 10.3390/v1010042. Epub 2009 Jun 8.
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.
本研究使用实时 PCR 检测法对小样本量进行了全面的 35 种呼吸道病原体筛查。初始热循环限制在 20 个循环，以避免试剂竞争，随后进行二次实时多重 PCR。需要补充半巢式人类偏肺病毒和小核糖核酸病毒 PCR 检测法来完成急性呼吸道病原体分析。在 121 例急性呼吸道症状儿童的鼻抽吸物中，有 85 例（70%）检测到潜在病原体。在这些样本中，有 29 例（24%）检测到多种病原体。串联多重实时 PCR 是一种快速检测多种病原体的有效方法。
