Chikungunya and dengue, two arboviral infections are common in South-East Asia and their early clinical manifestations are very similar hence it is important to discriminate between them as early as possible for better clinical management. The aim of this study was to design a rapid, sensitive and specific method for the differential diagnosis of these two viruses simultaneously. A rapid one-tube duplex RT-PCR assay was developed that requires 110 min including RNA extraction, RT-PCR and agarose gel electrophoresis by using a novel Taq polymerase with high processivity. This one-tube duplex RT-PCR system with primers designed from the conserved regions of the genome allowed discrimination between the two viral groups. Bioinformatics analysis of the DNA sequences from PCR amplified products confirmed that this method was very specific and accurate. The time required for this duplex RT-PCR was comparable to the standard IgM capture ELISA method. This novel approach would help to diagnose specifically and accurately these two closely related arboviruses and enable early detection from blood. This method could be applied in resource limited settings, for surveillance in endemic regions or for routine epidemiological screening.

译文

基孔肯雅热和登革热,两种虫媒病毒感染在东南亚很常见,它们的早期临床表现非常相似,因此尽早区分它们以更好的临床管理很重要。这项研究的目的是设计一种快速,灵敏和特异的方法来同时鉴别诊断疾病这两种病毒。开发了一种需要110分钟的快速单管双链rt-pcr分析,包括RNA提取、rt-pcr和琼脂糖凝胶电泳,通过使用具有高加工性的新型Taq聚合酶。这种单管双链rt-pcr系统具有从基因组保守区域设计的引物,可以区分两个病毒组。对PCR扩增产物DNA序列的生物信息学分析证实,该方法非常特异性和准确性。这种双重rt-pcr所需的时间与标准IgM捕获ELISA方法相当。这种新颖的方法将有助于特异性和准确地诊断这两种密切相关的虫媒病毒,并能够从血液中早期发现。此方法可用于资源有限的环境,流行地区的监测或常规流行病学筛查。

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