Development of DNA probes for the identification of sibling species A of the Anopheles culicifacies(Diptera Culicidae) Complex

Three highly repetitive DNA sequences Rp36, Rp217 and Rp234, have been isolated from Anopheles culicifacies Giles sensu lato . The cloned DNA sequences were found at a higher copy number in species B and C, than in species A of the A. culicifacies complex. These sequences may therefore be used as DNA probes to distinguish species A from the other two species, using a 200-fold dilution of a single mosquito DNA extract in a dot-blot hybridization assay. Rp36 and Rp217 have been completely sequenced. Internal repeats were absent in Rp36. Two related core sequences of 13 and 16 bp were found tandemly repeated in Rp217. These probes enable the rapid detection of species A of A. culicifacies in field investigations.