Ballinger-Crabtree M E, Black W C, Miller B R
Medical Entomology and Ecology Branch, National Center for Infectious Diseases, Centers for Disease Control, Fort Collins, Colorado.
Am J Trop Med Hyg. 1992 Dec;47(6):893-901. doi: 10.4269/ajtmh.1992.47.893.
Amplification of random regions of genomic DNA using 10-base primers in the random-amplified polymorphic DNA polymerase chain reaction (RAPD-PCR) was used to differentiate and identify mosquito populations based on genetic variation. Genomic DNA was extracted from individual mosquitoes from 11 geographic populations of Aedes aegypti and amplified in PCR reactions using single primers of arbitrary nucleotide sequence. Discriminant analysis of the population frequencies of RAPD fragments produced using three different primers allowed accurate discrimination between the geographic populations in 89% of individuals and between subspecies (Ae. aegypti aegypti versus Ae. aegypti formosus) in 100% of mosquitoes tested. The genetic relatedness of the populations was estimated using three different statistical methods, and unknown populations were correctly classified in a blind test. These results indicate that the RAPD-PCR technique will be useful in studies of arthropod molecular taxonomy and in epidemiologic studies of the relatedness of geographic populations and vector movement.
在随机扩增多态性DNA聚合酶链反应（RAPD-PCR）中，使用10碱基引物扩增基因组DNA的随机区域，以根据遗传变异来区分和鉴定蚊虫种群。从埃及伊蚊11个地理种群的单个蚊虫中提取基因组DNA，并使用任意核苷酸序列的单引物在PCR反应中进行扩增。对使用三种不同引物产生的RAPD片段的种群频率进行判别分析，在89%的个体中能够准确区分地理种群，在100%检测的蚊虫中能够区分亚种（埃及伊蚊埃及亚种与埃及伊蚊指名亚种）。使用三种不同的统计方法估计种群的遗传相关性，并在盲测中正确分类未知种群。这些结果表明，RAPD-PCR技术将有助于节肢动物分子分类学研究以及地理种群相关性和病媒移动的流行病学研究。
