Use of Genetic Polymorphisms Detected by the Random-Amplified Polymorphic DNA Polymerase Chain Reaction (RAPD-PCR) for Differentiation and Identification of Aedes aegypti Subspecies and Populations

Mary E. Ballinger-CrabtreeMedical Entomology and Ecology Branch, Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control, Department of Environmental Health, Colorado State University, Fort Collins, Colorado

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William C. Black IVMedical Entomology and Ecology Branch, Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control, Department of Environmental Health, Colorado State University, Fort Collins, Colorado

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Barry R. MillerMedical Entomology and Ecology Branch, Division of Vector-Borne Infectious Diseases, National Center for Infectious Diseases, Centers for Disease Control, Department of Environmental Health, Colorado State University, Fort Collins, Colorado

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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.

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