A method to increase efficiency in testing pooled field-collected mosquitoes
A method to increase efficiency in testing pooled field-collected mosquitoes
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DOI：
10.2987/5671.1
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发表时间：
2008-06-01
影响因子：
1
通讯作者：
Mores, Christopher N.
中科院分区：
文献类型：
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作者：
Chisenhall, Daniel M.;Vitek, Christopher J.;Mores, Christopher N.
测试现场捕获的蚊子集合可能会导致数千个池，并且每个测试50只蚊子的池既耗时又成本高昂。因此，我们已经开发出一种替代方法来测试蚊子池虫媒病毒，利用超级池策略。当处理蚊子样本以提取病毒RNA并随后通过定量实时聚合酶链反应进行病毒检测时，每个样本池都单独进行检测。使用本文所述的方法，将10个样本池各0.025 ml合并为一个超级样本池，用于RNA提取和检测。当发现病毒阳性超级合并液样本时，对构成该样本的原始10个合并液中的每一个进行单独检测，以发现特定的阳性样本。通过在初始超级池筛选后重新测试原始样本，我们仍然能够获得最小感染率或最大似然估计的可靠估计。为了测试这一原理，我们创建了已知滴度的受控蚊子池，并将它们置于我们的超级池过程中。我们能够检测到我们的整个范围的实验室创建的池作为西尼罗河病毒（WNV）阳性。2005年，我们实验室的实地监测工作对4，000多个蚊池进行了测试，其中8个样品呈WNV阳性。我们发现，所有这些现场样品都被检测为西尼罗河病毒阳性使用超级池方法，并包含计算的病毒滴度从
Testing field-caught mosquito collections can result in thousands of pools, and testing pools of 50 mosquitoes each can be both time consuming and cost prohibitive. Consequently, we have developed an alternative approach to testing mosquito pools for arboviruses, utilizing a superpool strategy. When mosquito samples are processed for extraction of viral RNA and subsequent virus testing via quantitative real-time polymerase chain reaction, each pool is tested individually. Using the method described here, 0.025 ml from each of 10 pools is combined into a superpool for RNA extraction and testing. When a virus-positive superpool sample is found, each of the original 10 pools that constitute this sample is tested individually in order to find the specific positive sample. By retesting the original samples after the initial superpool screen, we are still able to obtain reliable estimates for minimum infection rates or maximum likelihood estimations. To test this principle, we created controlled mosquito pools of known titer and subjected them to our superpool process. We were able to detect our entire range of laboratory-created pools as being West Nile virus (WNV) positive. In 2005, field Surveillance efforts from Our laboratory resulted in over 4,000 mosquito pools tested, with 8 resulting WNV-positive samples. We found that all of these field samples were detected as WNV positive using the superpool method and contained calculated virus titres from