通过实时TaqMan逆转录聚合酶链反应快速检测肠道病毒71型
Rapid detection of enterovirus 71 by real-time TaqMan RT-PCR.
作者信息
Tan Eng Lee, Yong Li Li Gaynor, Quak Seng Hock, Yeo Wei Cheng Andrea, Chow Vincent Tak Kwong, Poh Chit Laa
机构信息
Department of Microbiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore.
出版信息
J Clin Virol. 2008 Jun;42(2):203-6. doi: 10.1016/j.jcv.2008.01.001. Epub 2008 Mar 4.
BACKGROUND
Enterovirus 71 (EV71) is the main etiological agent of Hand, Foot and Mouth Disease (HFMD) and has been associated with neurological complications which resulted in fatalities during recent outbreaks in Asia Pacific region.
OBJECTIVE
Develop a real-time TaqMan RT-PCR for rapid detection of EV71.
STUDY DESIGN
Specific primers and probe were designed based on highly conserved VP1 region of EV71. The sensitivity of the real-time RT-PCR was evaluated with 67 clinical specimens collected from pediatric patients with suspected HFMD.
RESULTS
Our real-time TaqMan RT-PCR showed 100% specificity in detecting EV71 and showed an analytical sensitivity of 5 viral copies. High sensitivity was also achieved in detecting EV71 directly from clinical specimens.
CONCLUSIONS
Real-time TaqMan RT-PCR offers a rapid and sensitive method to detect EV71 from clinical specimens, and will allow quarantine measures to be taken more effectively during outbreaks.
背景
肠道病毒71型（EV71）是手足口病（HFMD）的主要病原体，在最近亚太地区的疫情爆发中，它与导致死亡的神经系统并发症有关。
目的
开发一种用于快速检测EV71的实时TaqMan RT-PCR方法。
研究设计
基于EV71高度保守的VP1区域设计特异性引物和探针。用从疑似手足口病的儿科患者中收集的67份临床标本评估实时RT-PCR的灵敏度。
结果
我们的实时TaqMan RT-PCR在检测EV71时显示出100%的特异性，分析灵敏度为5个病毒拷贝。直接从临床标本中检测EV71也具有高灵敏度。
结论
实时TaqMan RT-PCR提供了一种从临床标本中快速灵敏地检测EV71的方法，并将使在疫情爆发期间能更有效地采取检疫措施。
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