用于登革病毒RNA定量的定量竞争性逆转录-PCR
Quantitative competitive reverse transcription-PCR for quantification of dengue virus RNA.
作者信息
Wang W K, Lee C N, Kao C L, Lin Y L, King C C
机构信息
Institute of Microbiology, Taipei, Taiwan.
出版信息
J Clin Microbiol. 2000 Sep;38(9):3306-10. doi: 10.1128/JCM.38.9.3306-3310.2000.
Abstract
A quantitative competitive reverse transcription-PCR assay was developed to quantify dengue virus RNA in this study. The main features include a primer pair targeting a highly conserved region in the capsid and the addition of competing RNA that contains an internal deletion to provide a stringent internal control for quantification. It can be utilized to quantify RNA isolated from the four dengue virus serotypes but not RNA isolated from other flaviviruses, including Japanese encephalitis virus and hepatitis C virus, both prevalent in Asia. It can also be used to quantify dengue virus RNA isolated from the plasma of infected individuals. The sensitivity of the assay was estimated to be 10 to 50 copies of RNA per reaction, and twofold differences in virus titer are distinguishable. This assay is a convenient, sensitive, and accurate method for quantification and can be used to further understanding of the pathogenesis of dengue virus infection.
摘要
本研究开发了一种定量竞争性逆转录 - PCR 检测方法来定量登革病毒 RNA。其主要特点包括一对靶向衣壳中高度保守区域的引物对，以及添加含有内部缺失的竞争性 RNA，以提供严格的定量内部对照。它可用于定量从四种登革病毒血清型分离的 RNA，但不能用于定量从其他黄病毒分离的 RNA，包括在亚洲流行的日本脑炎病毒和丙型肝炎病毒。它还可用于定量从感染个体血浆中分离的登革病毒 RNA。该检测方法的灵敏度估计为每个反应 10 至 50 个 RNA 拷贝，病毒滴度的两倍差异是可区分的。该检测方法是一种方便、灵敏且准确的定量方法，可用于进一步了解登革病毒感染的发病机制。
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