Molecular diagnosis and analysis of Chikungunya virus
Molecular diagnosis and analysis of Chikungunya virus
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DOI：
10.1016/j.jcv.2007.05.008
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发表时间：
2007-08-01
影响因子：
8.8
通讯作者：
Lloyd, Graham
中科院分区：
文献类型：
--
作者：
Edwards, Carolyn J.;Welch, Stephen R.;Lloyd, Graham
背景：2005年3月，基孔肯雅热在印度洋岛屿爆发。在这些岛屿中，这种疾病的病例数急剧上升，影响了印度的100多万人。2006年，前往这些地区的旅行者已经返回英国，这种疾病导致基孔肯雅病毒（CHIKV）血清阳性样本的年数量增加了15倍以上。目的：针对CHIKV开发了实时RT-PCR检测，并设计用于检测当前流行的病毒株以及其他基因型。它的灵敏度进行了比较，与现有的标准RT-PCR检测和以前发表的实时assay.StudY设计：一个实时RT-PCR检测进行了优化和评价，使用一组55个临床血清样本和合成的RNA转录作为阳性对照。结果：实时荧光定量RT-PCR的灵敏度是常规阻断RT-PCR的10倍，最低可检测到20个拷贝的RNA转录本。当与另一种公开的TaqMan测定相比时，该测定在检测病毒的爆发株中也具有10倍的提高的灵敏度。对来自印度、毛里求斯和塞舌尔的CHIKV患者的基因序列进行分析，结果显示与来自印度洋R6 union岛的已发表序列具有高度的相似性。结论：建立了一种灵敏、快速的CHIKV实时荧光定量PCR检测方法。(c)2007 Elsevier B. V.保留所有权利。
Background: In March 2005 a Chikungunya fever outbreak began on the islands of the Indian Ocean. The number of cases of this disease dramatically rose amongst these islands before affecting over a million people in India. Travellers to these regions have returned to the UK with the disease leading to a greater than 15-fold increase in the annual number of Chikungunya virus (CHIKV) sero-positive samples in 2006.Objectives: A real-time RT-PCR test was developed for CHIKV and designed to detect currently circulating strains of virus as well as other genotypes. Its sensitivity was compared with an existing standard RT-PCR assay and a previously published real-time assay.StudY design: A real-time RT-PCR assay was optimised and evaluated using a panel of 55 clinical serum samples and a synthetic RNA transcript as a positive control. Nucleotide sequencing of part of the El gene of CHIKV was used to investigate the relatedness of the samples.Results: The real-time RT-PCR was I 0-fold more sensitive than a conventional block-based RT-PCR and could detect as low as 20 copies of RNA transcript. The assay also had I 0-fold improved sensitivity in detecting the outbreak strain of virus when compared to another published TaqMan assay. Analysis of sequences from patients that had travelled to India, Mauritius or the Seychelles showed high similarity with published sequences from the Indian Ocean island of R6union.Conclusions: A sensitive and rapid real-time RT-PCR assay has been developed for CHIKV and tested against current isolates. (c) 2007 Elsevier B.V. All rights reserved.