A new quantitative RT-PCR method for sensitive detection of dengue virus in serum samples
A new quantitative RT-PCR method for sensitive detection of dengue virus in serum samples
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DOI：
10.1016/j.jviromet.2008.06.023
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发表时间：
2008-10-01
影响因子：
3.1
通讯作者：
Vassilev, Ventzislav
中科院分区：
文献类型：
--
作者：
Sadon, Nadine;Delers, Anne;Vassilev, Ventzislav
为了检测和鉴定血清样本中的登革热血清型，我们开发了一个单步定量逆转录-聚合酶链反应（Q-RT-PCR）检测（简称Q-PCR）。引物组选自病毒基因组的衣壳区。使用特异性引物和荧光TaqManTM探针在两个单独的双链扩增反应中检测登革热血清型1/3和2/4。使用从泰国曼谷的患者中收集的97份代表性人血清的样本组，比较了用该Q-PCR和经典巢式RT-PCR（N-PCR）测定法获得的结果。结果表明，Q-PCR是一种快速，灵敏和可重复的工具，用于检测和定量的四个登革热血清型的临床样本，因此非常感兴趣的诊断用途或大型队列研究。(C)2008 Elsevier B. V.保留所有权利。
In order to detect and identify dengue serotypes in serum samples, we developed a single-step quantitative reverse transcription-polymerase chain reaction (Q-RT-PCR) assay (referred to as Q-PCR). Sets of primers were selected from the capsid region of the viral genome. Dengue serotypes 1/3 and 2/4 were detected in two separate duplex amplification reactions using specific primers and fluorogenic TaqMan (TM) probes. Results obtained with this Q-PCR and the classical nested RT-PCR (N-PCR) assays were compared using a panel of 97 representative human sera collected from patients in Bangkok, Thailand. It is shown that the Q-PCR is a rapid, sensitive and reproducible tool for the detection and quantitation of the four dengue serotypes in clinical samples, and therefore of great interest for diagnostic use or for large cohort studies. (C) 2008 Elsevier B.V. All rights reserved.