PCR and dissection as tools to monitor filarial infection of Aedes polynesiensis mosquitoes in French Polynesia.
PCR and dissection as tools to monitor filarial infection of Aedes polynesiensis mosquitoes in French Polynesia.
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PCR和解剖作为监测法氏多尼西斯蚊子的丝状感染的工具。
DOI：
10.1186/1475-2883-5-2
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发表时间：
2006-02-24
期刊：
影响因子：
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通讯作者：
Legrand, Anne-Marie
中科院分区：
文献类型：
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作者：
Plichart, Catherine;Sechan, Yves;Davies, Neil;Legrand, Anne-Marie
昆虫学方法可能为监测法属波利尼西亚丝虫病的传播提供重要工具。为了标准化我们的PCR方法和完善我们的协议，以评估丝虫感染水平的蚊子，我们比较解剖的载体，玻里尼西亚伊蚊，池筛选聚合酶链反应（PS-PCR）测定。(1)在法属波利尼西亚莫雷阿岛的五个地区收集了人类登陆捕获的蚊子。(2)将捕获的一部分蚊子解剖以获得班氏吴策线虫幼虫。(3)重复测试人工饲养的蚊子（未感染的以及实验感染的蚊子）以优化PS-PCR方案（用内部标准化系统和特异于Ssp 1重复序列的引物测试来自1-50只合并蚊子的DNA提取物。通过凝胶电泳分析PCR产物）。(4)根据优化的方案，通过PS-PCR检测捕获的蚊子的另一部分。野外蚊虫感染W。通过解剖（L1-L3），班氏病的范围为1%-8%，通过PS-PCR测定，感染流行率的点估计值范围为0.4%-3.7%。通过解剖和PCR确定的幼虫感染率之间存在中等强度的相关性。结果表明，PS-PCR方法特异性强，灵敏度高。我们通过解剖蚊子获得了类似但不完全相同的结果。PS-PCR似乎足以在消除丝虫病计划的背景下检测大量蚊子。本文讨论了应用昆虫学方法监测丝虫病的作用和优点。
Entomological methods may provide important tools for monitoring the transmission of filariasis in French Polynesia. In order to standardize our PCR method and refine our protocol to assess filarial infection levels in mosquitoes, we compared dissection of the vector, Aedes polynesiensis, with the poolscreening polymerase chain reaction (PS-PCR) assay. (1) Mosquitoes were collected in human landing catches in five areas in Moorea island, French Polynesia. (2) A fraction of the captured mosquitoes was dissected for Wuchereria bancrofti larvae. (3) Laboratory-reared mosquitoes (uninfected as well as experimentally infected ones) were repeatedly tested to optimize a PS-PCR protocol (DNA extracts from 1–50 pooled mosquitoes were tested with an internal standardized system and primers specific for the Ssp1 repeat sequence. PCR products were analysed by gel electrophoresis). (4) Another fraction of the captured mosquitoes was assayed by PS-PCR according the optimized protocol. The prevalence of field-mosquito infection with W. bancrofti ranged from 1 to 8 % by dissection (L1–L3) and point estimates of infection prevalence, as assayed by PS-PCR, ranged from 0.4 to 3.7 %. There was a moderately strong correlation between larval infection rates as determined by dissection and PCR. Our results suggest that the PS-PCR assay is specific and highly sensitive for detecting parasite DNA. We obtained similar although not identical results with dissections of mosquitoes. PS-PCR appears to be adequate for testing large numbers of mosquitoes in the context of filariasis elimination programs. The role and advantages of using entomologic methods to monitor filariasis programs are discussed.