Loop-mediated isothermal amplification applied to filarial parasites detection in the mosquito vectors: Dirofilaria immitis as a study model.
Loop-mediated isothermal amplification applied to filarial parasites detection in the mosquito vectors: Dirofilaria immitis as a study model.
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DOI：
10.1186/1756-3305-2-15
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发表时间：
2009-03-15
影响因子：
3.2
通讯作者：
Kanuka H
中科院分区：
文献类型：
--
作者：
Aonuma H;Yoshimura A;Perera N;Shinzawa N;Bando H;Oshiro S;Nelson B;Fukumoto S;Kanuka H
尽管我们最近在了解节肢动物病媒传播人畜共患传染病背后的基本生物学方面取得了进展，但这些疾病仍然威胁着公共卫生问题。为了有效控制病媒和治疗，精确抽样以显示此类疾病的流行情况是至关重要的。为了建立一种快速、简便的检测人畜共患病原体传播媒介的方法，应用环介导等温扩增(LAMP)技术，以埃及伊蚊和犬心虫为研究对象，建立了丝虫传播实验模型。LAMP反应扩增细胞色素氧化酶亚基I基因，当检测到单个纯化的微丝虫时，显示出高灵敏度。重要的是，LAMP反应的稳健性是在识别出只携带一种寄生虫的受感染蚊子时揭示的，这一水平很容易被传统的显微镜分析忽视。此外，在野外捕获的蚊子中成功检测到蠕形虫，证明了它在现场调查中的适用性。LAMP方法简便、灵敏、可靠，可作为蚊媒携带丝虫寄生虫的常规诊断方法。这种方法不仅可以用于犬丝虫病的调查，还可以用于淋巴丝虫病的调查，这是另一个主要的公共卫生问题。因此，该方法为丝虫病流行地区丝虫寄生虫的检测提供了一种有用的诊断方法。
Despite recent advances in our understanding of the basic biology behind transmission of zoonotic infectious diseases harbored by arthropod vectors these diseases remain threatening public health concerns. For effective control of vector and treatment, precise sampling indicating the prevalence of such diseases is essential. With an aim to develop a quick and simple method to survey zoonotic pathogen-transmitting vectors, LAMP (loop-mediated isothermal amplification) was applied to the detection of filarial parasites using a filarial parasite-transmitting experimental model that included one of the mosquito vectors, Aedes aegypti, and the canine heartworm, Dirofilaria immitis. LAMP reactions amplifying the cytochrome oxidase subunit I gene demonstrated high sensitivity when a single purified D. immitis microfilaria was detected. Importantly, the robustness of the LAMP reaction was revealed upon identification of an infected mosquito carrying just a single parasite, a level easily overlooked using conventional microscopic analysis. Furthermore, successful detection of D. immitis in wild-caught mosquitoes demonstrated its applicability to field surveys. Due to its simplicity, sensitivity, and reliability, LAMP is suggested as an appropriate diagnostic method for routine diagnosis of mosquito vectors carrying filarial parasites. This method can be applied to the survey of not only canine filariasis but also lymphatic filariasis, another major public health problem. Therefore, this method offers great promise as a useful diagnostic method for filarial parasite detection in endemic filariasis regions.
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影响因子：
9.4
作者：
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