Identification of bloodmeals in haematophagous Diptera by cytochrome B heteroduplex analysis
Identification of bloodmeals in haematophagous Diptera by cytochrome B heteroduplex analysis
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DOI：
10.1046/j.1365-2915.1999.00193.x
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发表时间：
1999-07-01
影响因子：
1.9
通讯作者：
Unnasch, TR
中科院分区：
文献类型：
--
作者：
Boakye, DA;Tang, J;Unnasch, TR
我们开发了一种用于食血昆虫血粉鉴定的DNA检测方法。通过PCR扩增特异性宿主细胞色素B基因序列，并根据其在异双工试验中的迁移性进行分类。在黑蝇（双翅目：黑蝇科）中，人类细胞色素B DNA序列在摄入血粉3天后仍可被识别。在象牙海岸采集的舌蝇（双翅目：舌蝇科）中，根据其异双工模式和DNA序列确定血食来自家猪。显然，细胞色素B序列表现出足够的种间差异来区分哺乳动物宿主样本，而表现出最小的种内差异。在吸血昆虫食用后的数天内，血肉中DNA的稳定性使PCR-HDA测定能够可靠地用于宿主鉴定。
We developed a DNA assay for bloodmeal identification in haematophagous insects. Specific host cytochrome B gene sequences were amplified by PCR and classified on the basis of their mobility in a heteroduplex assay. In the blackfly Simulium damnosum s.l. (Diptera: Simuliidae), human cytochrome B DNA sequences were identifiable up to 3 days following ingestion of the bloodmeal. In the tsetse Glossina palpalis (Diptera: Glossinidae) collected from tsetse traps in Ivory Coast, bloodmeals were identified as taken from domestic pigs on the basis of their heteroduplex pattern and DNA sequence. Evidently the cytochrome B sequence shows sufficient interspecific variation to distinguish between mammalian host samples, while exhibiting minimal intraspecific variation. The stability of DNA in bloodmeals, for several days post-ingestion by haematophagous insects, allows PCR-HDA assays to be used reliably for host identification.