Rapid identification of virus-carrying mosquitoes using reverse transcription-loop-mediated isothermal amplification
Rapid identification of virus-carrying mosquitoes using reverse transcription-loop-mediated isothermal amplification
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DOI：
10.1016/j.jviromet.2008.10.023
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发表时间：
2009-03-01
影响因子：
3.1
通讯作者：
Kanuka, Hirotaka
中科院分区：
文献类型：
--
作者：
Perera, Namal;Aonuma, Hiroka;Kanuka, Hirotaka
蚊子是许多虫媒病毒传播周期中的重要媒介。考虑到全世界虫媒病毒感染的发病率不断增加，监测虫媒病毒的存在可被视为对人类和动物进行风险评估的重要初步步骤。为了满足这一需要，已加大努力开发快速可靠的诊断技术；以禽舍病毒（FHV）为模型，建立了一种单步逆转录环介导等温扩增（RT-LAMP）方法检测埃及伊蚊（Aedes aegypti）中的病毒。RT-LAMP反应的稳健性表明，它能够在30分钟内用整个蚊子体从“一体化”模板中检测出FHV。此外，RT-LAMP成功地鉴定了一只携带单个FHV颗粒的蚊子，这在传统分析（如斑块形成分析）中很容易被忽视。这些观察结果表明，在西尼罗热或登革热等病媒传播疾病普遍流行的地区，RT-LAMP对于病媒蚊子的常规诊断更为可靠和有用。(c) 2008 Elsevier B.V.版权所有
Mosquitoes are critical vectors in many arboviral transmission cycles. Considering the increasing incidence of arboviral infections throughout the world, monitoring of vector populations for the presence of an arbovirus could be considered an important initial step of risk assessment to humans and animals. In response to this need, increased efforts to develop rapid and reliable diagnostic techniques have been undertaken; a single-step reverse transcription-loop-mediated isothermal amplification (RT-LAMP) assay was developed to detect virus in vector mosquitoes (Aedes aegypti) using the Flock House Virus (FHV) as a model. The robustness of the RT-LAMP reaction was revealed by its ability to detect FHV from an "all-in-one" template using whole mosquito bodies within 30 min. Furthermore, RT-LAMP identified successfully a mosquito carrying just a single FHV particle, a level easily overlooked in conventional analysis such as plaque forming assays. These observations suggest that RT-LAMP is more reliable and useful for routine diagnosis of vector mosquitoes in regions where the prevalence of vector-borne diseases such as West Nile fever or dengue fever are common. (c) 2008 Elsevier B.V. All rights reserved.