Molecular differentiation of colonized human malaria vectors by 28S ribosomal DNA polymorphisms

Anopheles gambiae s.s. Giles, An. stephensi Liston, An. freeborni Aitken, and An. quadrimaculatus Say are cultured and studied in molecular genetic and transgenic laboratories with increasing frequency. With limited research space, these mosquitoes are often maintained in the same insectary. Under these conditions, cross-contamination of colonies can occur and have devastating consequences to affected research programs. We have developed a polymerase chain reaction-based assay targeting the 28S large subunit ribosomal RNA gene to easily differentiate between these four taxa and An. funestus Giles, which occurs in sympatry with An. gambiae. The resulting assay identifies individual mosquito preparations as well as all taxa within a mixed or pooled DNA template preparation.