Nucleic acid sequence-based amplification assays for rapid detection of West Nile and St. Louis encephalitis viruses
Nucleic acid sequence-based amplification assays for rapid detection of West Nile and St. Louis encephalitis viruses
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DOI：
10.1128/jcm.39.12.4506-4513.2001
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发表时间：
2001-12-01
影响因子：
9.4
通讯作者：
Kerst, AJ
中科院分区：
文献类型：
--
作者：
Lanciotti, RS;Kerst, AJ
本文报道了西尼罗病毒（WN）和圣路易斯脑炎病毒（SLE）核酸序列扩增（NASBA）检测方法的发展和应用。为NASBA检测开发了两种独特的检测格式：使用病毒特异性内部捕获探针和电化学发光的扩增后检测步骤（NASBA- ecl检测）和使用6-羧基荧光素标记的病毒特异性分子信标探针的实时检测（NASBA-信标检测）。将这些NASBA检测方法的敏感性和特异性与新描述的标准逆转录(RT)-PCR和系统性红斑狼疮病毒TaqMan检测方法以及先前发表的系统性红斑狼疮病毒TaqMan检测方法进行了比较。与病毒分离、TaqMan检测和标准RT-PCR相比，NASBA检测显示出卓越的敏感性和特异性，NASBA信标检测在不到1小时的时间内产生结果。这些检测应该在诊断实验室中实用，以补充现有的诊断检测方法，并作为在美国进行黄病毒监测的工具。
The development and application of nucleic acid sequence-based amplification (NASBA) assays for the detection of West Nile (WN) and St. Louis encephalitis (SLE) viruses are reported. Two unique detection formats were developed for the NASBA assays: a postamplification detection step with a virus-specific internal capture probe and electrochemiluminescence (NASBA-ECL assay) and a real-time assay with 6-carboxyfluorescein-labeled virus-specific molecular beacon probes (NASBA-beacon assay). The sensitivities and specificities of these NASBA assays were compared to those of a newly described standard reverse transcription (RT)-PCR and TaqMan assays for SLE virus and to a previously published TaqMan assay for WN virus. The NASBA assays demonstrated exceptional sensitivities and specificities compared to those of virus isolation, the TaqMan assays, and standard RT-PCR, with the NASBA-beacon assay yielding results in less than 1 h. These assays should be of utility in the diagnostic laboratory to complement existing diagnostic testing methodologies and as a tool in conducting flavivirus surveillance in the United States.