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首页> 外文期刊>Journal of Medical Entomology >Laboratory and field evaluation of polymerase chain reaction-based forensic DNA profiling for use in identification of human blood meal sources of Aedes aegypti (diptera: culicidae)
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Laboratory and field evaluation of polymerase chain reaction-based forensic DNA profiling for use in identification of human blood meal sources of Aedes aegypti (diptera: culicidae)

机译：基于聚合酶链反应的法医DNA分析的实验室和现场评估，用于鉴定埃及伊蚊的人血粉来源（双翅目：库蚊）

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We modified polymerase chain reaction (PCR)-based forensic DNA profiling for field studies on the feeding behavior of Aedes aegypti, the principal mosquito vector of dengue virus. Human DNA was extracted from oral swabs of human subjects and from blood-engorged mosquitoes, DNA was quantified by slot blot, and alleles at variable number tandem repeats and three short tandem repeats loci were amplified by PCR. Alleles were separated electrophoretically and then visualized by silver staining, A custom software program was written to match DNA fingerprints of potential human hosts to allelic profiles detected in engorged mosquitoes, and to calculate error rates for identification of human hosts of single and multiple-host blood meals. At 29~C in the laboratory, human DNA recovered from mosquito blood meals declined an average of 67% 8 h after feeding and 90% after 24 h. We obtained complete allelic profiles from seven of 10 mosquitoes collected after 24 h. In a field trial, complete DNA profiles were obtained successfully for 43 people living in a rural village in south central Thailand and for 20 of 100 Ae. aegypti that contained blood and were collected in those peoples' homes. Blood imbibed from more than one person was detected in 45% (9 of 20) of the meals. Sixty-five percent of the meals contained blood from nonresidents of the house in which the mosquito was collected or from people who were not profiled; data consistent with the hypothesis that human movement is important for the spread of dengue virus within and among communities. When using alleles at four loci, all of the Thais and nine members spanning three generations of a Chinese-American family had unique allelic profiles. Error rates from classifying possible multiplehost meals as single-host meals were low (1-8%), with the highest error associated with closely related people. Results from our laboratory and field studies indicated that DNA profiling can be used to study the details and epidemiological implications of Ae. aegypti blood-feeding behavior.
机译：我们修改了基于聚合酶链反应（PCR）的法医DNA谱图，用于对埃及伊蚊（登革热病毒的主要灭蚊载体）的摄食行为进行现场研究。从人类受试者的口腔拭子和血液充血的蚊子中提取人类DNA，通过狭缝印迹对DNA进行定量，并通过PCR扩增可变数目的串联重复和三个短串联重复位点的等位基因。电泳分离等位基因，然后通过银染可视化。编写了一个定制软件程序，以将潜在人类宿主的DNA指纹与饱食蚊子中检测到的等位基因谱进行匹配，并计算错误率以鉴定单宿主和多宿主血液的人类宿主饭菜。在实验室温度为29°C时，从蚊子血粉中回收的人类DNA在进食后8小时平均下降67％，在24小时后平均下降90％。我们从24小时后收集的10个蚊子中的7个获得了完整的等位基因图谱。在现场试验中，成功地获得了居住在泰国中南部一个乡村中的43个人和100 Ae中的20个人的完整DNA图谱。埃及血统，这些血统收集在这些人的家中。膳食中有45％（20人中有9人）检测到从一个以上的人身上吸收的血液。 65％的膳食中所含血液来自收集蚊子的房屋的非居民或未进行简介的人;数据与以下假设相符：人的流动对于登革热病毒在社区内部和社区之间的传播很重要。在四个位点使用等位基因时，所有泰国人和跨越华裔美国家庭三代的九个成员均具有独特的等位基因特征。将可能的多宿主膳食分类为单宿主膳食的错误率很低（1-8％），与紧密相关的人相关的错误率最高。我们实验室和现场研究的结果表明，DNA分析可用于研究Ae的细节和流行病学意义。埃及人的采血行为。

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