A Simplified Non Radioactive DNA Probe Technique for the Field Detection of Sibling Species A of the Anopheles culicifacies (Diptera: Culicidae) Complex

Abstract

Three cloned highly repetitive DNA sequences Rp36, Rp2l7 and Rp234 isolated from An. culicifacies Giles, sensu lato were developed as non radioactive DNA probes by using a biotinylated labeling and colorimetric detection system. These non radioactive DNA probes distinguish sibling species A from species Band C of the An. culicifacies complex in a dot blot hybridization assay using single mosquito DNA extracts diluted 50 fold. The biotinylated Rp217 probe was further assayed in a more simple procedure which involves the hybridization of blots prepared from squashed mosquito heads. This technique avoids the separate extraction of mosquito DNA and facilitates a number of samples to be processed rapidly while also allowing several field analyses to be carried out on one mosquito specimen.