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实时逆转录环介导等温扩增法快速鉴定基孔肯雅热和登革热病毒。

Rapid identification of Chikungunya and Dengue virus by a real-time reverse transcription-loop-mediated isothermal amplification method.

机构信息

Department of Respiratory, Tangdu Hospital, Fourth Military Medical University, Xi'an, China.

出版信息

Am J Trop Med Hyg. 2012 Nov;87(5):947-53. doi: 10.4269/ajtmh.2012.11-0721. Epub 2012 Sep 10.

Abstract

Both Chikungunya and Dengue virus belong to the acute arthropod-borne viruses. Because of the lack of specific symptoms, it is difficult to distinguish the two infections based on clinical manifestations. To identify and quantitatively detect Chikungunya and Dengue viruses, a real-time accelerated reverse-transcription-loop-mediated isothermal amplification (RT-LAMP) platform was developed, and 26-confirmed RNA samples, 42 suspects, and 18 healthy serum samples were evaluated by the method. The RT-polymerase chain reaction (PCR) and cDNA sequencing were used as references. The results showed that it could identify the Chikungunya and Dengue virus RNA correctly in all antibody-positive samples within 1 hour, without any cross-reactions. The virus load of the positive samples was quantitatively detected with a turbidimeter. The sensitivity was 100% and specificity was 95.25%. The findings indicate that the RT-LAMP is an effective method for rapid quantity detection of Chikungunya virus and Dengue virus in serum samples with convenient operation, high specificity, and high sensitivity.

摘要

基孔肯雅热病毒和登革热病毒均属于急性虫媒病毒。由于缺乏特异性症状,临床上难以区分这两种感染。为了鉴定和定量检测基孔肯雅热病毒和登革热病毒,开发了一种实时加速逆转录环介导等温扩增(RT-LAMP)平台,并使用该方法评估了 26 份确诊 RNA 样本、42 份疑似样本和 18 份健康血清样本。将 RT-聚合酶链反应(PCR)和 cDNA 测序作为参考。结果表明,该方法可在 1 小时内正确识别所有抗体阳性样本中的基孔肯雅热病毒和登革热病毒 RNA,无任何交叉反应。用浊度计定量检测阳性样本中的病毒载量。灵敏度为 100%,特异性为 95.25%。研究结果表明,RT-LAMP 是一种快速定量检测血清样本中基孔肯雅热病毒和登革热病毒的有效方法,操作方便,特异性和灵敏度高。

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