Detection of North American eastern and western equine encephalitis viruses by nucleic acid amplification assays
Detection of North American eastern and western equine encephalitis viruses by nucleic acid amplification assays
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DOI：
10.1128/jcm.41.1.379-385.2003
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发表时间：
2003-01-01
影响因子：
9.4
通讯作者：
Lanciotti, RS
中科院分区：
文献类型：
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作者：
Lambert, AJ;Martin, DA;Lanciotti, RS
我们已经开发了基于核酸序列的扩增（NASBA），标准逆转录PCR（RT-PCR），和TaqMan核酸扩增检测北美东部马脑炎（EAE）和西部马脑炎（WEE）病毒RNA从现场和临床样品中收集的样品的快速检测。这些试验的灵敏度进行了比较，病毒分离。虽然所有三种类型的核酸扩增测定都提供了与Vero细胞中病毒分离相当的病毒RNA的快速检测，但用于北美猪瘟和WEE病毒RNA的TaqMan测定是最灵敏的。我们已经证明这些检测是特定的北美ESTA和WEE病毒RNA通过测试地理和时间上不同的菌株ESTA和WEE病毒沿着电池的相关和不相关的节肢动物传播的病毒。此外，所有三种类型的核酸扩增测定已被用于检测蚊子和脊椎动物组织样品中的北美EAE和WEE病毒RNA。核酸扩增的灵敏度、特异性和快速性证明了NASBA、标准RT-PCR和TaqMan测定法在研究和诊断环境中检测北美猪瘟和WEE病毒RNA的有用性。
We have developed nucleic acid sequence-based amplification (NASBA), standard reverse transcription PCR (RT-PCR), and TaqMan nucleic acid amplification assays for the rapid detection of North American eastern equine encephalitis (EEE) and western equine encephalitis (WEE) viral RNAs from samples collected in the field and clinical samples. The sensitivities of these assays have been compared to that of virus isolation. While all three types of nucleic acid amplification assays provide rapid detection of viral RNAs comparable to the isolation of viruses in Vero cells, the TaqMan assays for North American EEE and WEE viral RNAs are the most sensitive. We have shown these assays to be specific for North American EEE and WEE viral RNAs by testing geographically and temporally distinct strains of EEE and WEE viruses along with a battery of related and unrelated arthropodborne viruses. In addition, all three types of nucleic acid amplification assays have been used to detect North American EEE and WEE viral RNAs from mosquito and vertebrate tissue samples. The sensitivity, specificity, and rapidity of nucleic acid amplification demonstrate the usefulness of NASBA, standard RT-PCR, and TaqMan assays, in both research and diagnostic settings, to detect North American EEE and WEE viral RNAs.