Anopheles fluviatilis is one of the major vectors of malaria in India and is a complex of at least three cryptic species (Species S, T and U). Identification of cryptic species of An. fluviatilis complex is of paramount interest owing to differences in their vectorial efficiency and preference to feed on man. These members are morphologically indistinguishable at any stage of life cycle and can be identified only by the examination of species-specific inversions on the polytene chromosomes. We report allele specific polymerase chain reaction (ASPCR)-based diagnostic method for the differentiation of all members of An. fluviatilis complex that is applicable to all stages and sexes of the mosquito. For the development of diagnostic assay, D3 domain of 28S ribosomal DNA of all the three members was amplified, sequenced, and analyzed for species-specific differences in nucleotide sequences. Primers designed based on these differences were used for allele- specific amplification to differentiate the members. The PCR assay was evaluated against cytologically confirmed field-collected individuals from various parts of India representing different sympatric associations
Species 1: Diptera Culicidae Anopheles fluviatilis
Keywords: polymerase chain reaction
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