Abstract: Objective To study the sensitive of TaqMan MGB Real-time polymerase chain reaction(PCR) and nested RT-PCR on detecting Dengue virus(DV) from Aedes albopictus and establish a sensitive, specific, and repetitive method for DV diagnosis. Methods Females adults were artificially infected with Den-2 in lab, different infected mosquito concentration (mosquitos/1000 μl) and 50 mosquitos/pool were designed and processed for virus detection by TaqMan MGB Real-time PCR and nested RT-PCR. Fluorescence signal and electrophoresis showed the results. Results For all pool, the lightest concentration that can be detected was 3 mosquitos/1000 μl for two-step TaqMan MGB Real-time PCR, and 5 mosquitos/1000 μl for one-step TaqMan MGB Real-time PCR and nested RT-PCR. Conclusion Two-step TaqMan MGB is more sensitive, specific, scientific and rapid to detect DV than nested RT-PCR, and is a good surveillance method for DV in A.albopictus.

摘要： 目的 比较TaqMan MGB探针实时聚合酶链反应(TaqMan MGB Real-time PCR)和巢式反转录聚合酶链反应(巢式RT-PCR)检测白纹伊蚊体内登革热病毒(Dengue virus,DV)的敏感性差异,建立一种敏感、特异、重复性好的DV鉴定方法。方法 在实验室使雌性白纹伊蚊人工感染Den-2,在50只/组情况下,设不同的感染蚊虫浓度(只/1000μl),利用TaqMan MGB Real-time PCR和巢式RT-PCR检测,根据荧光信号和电泳判断结果。结果 二步法TaqMan MGB Real-ti me PCR可检测到标本中感染蚊虫的最低浓度为3只/1000μl,一步法TaqMan MGB Real-ti me PCR和巢式RT-PCR可检测到标本中感染蚊虫的最低浓度为5只/1000μl。结论 二步法TaqMan MGB探针检测白纹伊蚊体内的DV敏感度高,特异性好,快速、科学,是白纹伊蚊携带DV指数较理想的监测方法。

关键词: 白纹伊蚊, 登革热2型病毒, TaqMan MGB探针, 实时聚合酶链反应, 巢式反转录聚合酶链反应