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

Abstract

Three highly repetitive DNA sequences, Rp36, Rp217 and Rp234, were isolated from A. culicifacies s.l. 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 2 species, using a 200-fold dilution of a single mosquito DNA extract in a dot-blot hybridization assay. Rp36 and Rp217 were completely sequenced. Internal repeats were absent in Rp36. Two related core sequences of 134 and 16 bp were found tandemly repeated in Rp217. These probes enable the rapid detection of species A of A. culicifacies in field investigations