A simplified non radioactive DNA probe technique for the field detection of sibling species A of the Anopheles culicifacies (Diptera: Culicidae) complex

B.G.D.N.K. De Silva, G. P. G. M. D. Hapugoda, E. H. Karuuanayake

Abstract


Three cloned highly repetitive DNA sequences Rp36, Rp217 and Rp234 isolated from An.culicifacies Giles, sensu lato were developed as non radioactive DNA probes by using a biotinylatedlabeling and colorimetric detection system. These non radioactive DNA probes distinguish siblingspecies A from species B and C of the An. culicifacies complex in a dot blot hybridization assayusing single mosquito DNA extracts diluted 50 fold. The biotinylated Rp217 probe was furtherassayed in a more simple procedure which involves the hybridization of blots prepared fromsquashed mosquito heads. This technique avoids the separate extraction of mosquito DNA andfacilitates a number of samples to be processed rapidly while also allowing several field analyses tobe carried out on one mosquito specimen.

Key words : Anopheles culicifacies, non radioactive DNA probes


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