Simple nonradioactive DNA hybridization method for identification of sibling species of Anopheles dirus (Diptera: Culicidae) complex.

A simple method was developed for species identification of mosquitoes in the Anopheles dirus complex found in Thailand using horseradish peroxidase-labeled DNA probes and a chemiluminescent detection system. Species-specific DNA probes for Anopheles dirus, species B, C, and D detected 1-5 ng of target DNA, a sensitivity that was comparable with the radioisotopic detection system. Identification of individual mosquitoes was performed by dot-blot analysis of crude mosquito DNA. The method allowed identification using third or fourth instars as well as the adult head, thorax, or abdomen. This technique successfully identified the sibling species of the A. dirus complex in field collections.