Identification of mammalian blood meals in mosquitoes by a multiplexed polymerase chain reaction targeting cytochrome b
Identification of mammalian blood meals in mosquitoes by a multiplexed polymerase chain reaction targeting cytochrome b
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DOI：
10.4269/ajtmh.2005.73.336
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发表时间：
2005-08-01
影响因子：
3.3
通讯作者：
Norris, DE
中科院分区：
文献类型：
--
作者：
Kent, RJ;Norris, DE
迄今为止，还没有聚合酶链反应诊断技术能够通过琼脂糖凝胶电泳后的 DNA 片段大小直接识别来自蚊子的哺乳动物血粉。我们开发了一种基于线粒体细胞色素 b 的脊椎动物特异性多重引物组，用于识别现场采集的蚊子的哺乳动物血液宿主。尽管该工具是为研究非洲疟疾媒介而设计的，但该工具的应用并不限于该疾病系统。这种诊断技术对在赞比亚和马里收集的干按蚊和库西尼野外标本进行的验证表明，可以在采集后 2-7 个月识别出血粉。时程实验表明，在喂食后 24-30 小时的冷冻蚊子腹部中可以检测到宿主 DNA。此外，可以在一只蚊子中检测到来自不同哺乳动物的多种血粉。这种诊断分析将成为一种有价值的工具，用于识别人类和其他哺乳动物宿主存在的现场采集的蚊子的血粉。
To date, no polymerase chain reaction diagnostic technique exists to directly identify mammalian blood meals from mosquitoes by sized DNA fragments following agarose gel electrophoresis. We have developed a vertebrate-specific multiplexed primer set based on mitochondrial cytochrome b to identify the mammalian blood hosts of field-collected mosquitoes. Although designed for the study of African malaria vectors, the application of this tool is not restricted to this disease system. Validation of this diagnostic technique on dried anopheline and culicine field specimens collected in Zambia and Mali demonstrated that blood meals could be identified 2-7 months after collection. Time course experiments showed that host DNA was detectable in frozen mosquito abdomens 24-30 hours post-feeding. Additionally, multiple blood meals from different mammals could be detected in a single mosquito. This diagnostic assay will be a valuable tool for identifying the blood meals of field-collected mosquitoes where people and alternative mammal hosts are present.