[PCR-RFLP and sequencing studies of malaria vectors (Diptera, Culicidae, Anopheles) in Kyrgyzstan].

ABSTRACT

The results of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, obtained after treatment of PCR-products with restriction endonuclease CfoI, could identify two members of the Anopheles maculipennis complex An. maculipennis and An. artemievi. Treatment of amplification products with restriction endonuclease BsuI gave rise to fragment lengths of 192 and 218 bp, characteristic of An. artemievi, in the populations of the Talas (settlement of Kizil-Adyr, Kara-Bura District), Dzhelalabad (towns of Tashkumyr and Kara-Kul), and Osh (town of Gulcha, Alai District; village of Langar, Kara-Suisky District) Regions. After treatment of PCR-products with restriction endonuclease BstACI, fragment lengths of 292 and 150 bp, characteristic of An. messeae, were obtained for the mosquitoes of Issyk-Kul (town of Balykchi) and Naryn (settlement of Kochkorka, Kochkor District) Regions. To identify the molecular forms of An. superpictus, the investigators sequenced the amplification products obtained by PCR with 5.8S and 28S rRNA gene-specific primers. Analysis of the primary structure of the second internal transcribed spacer, by using the international databases, has indicated that molecular form X is prevalent in the study districts of Kyrgyzstan. The COI-COII region of the mitochondrial genome of the vector also underwent PCR-RFLP analysis. Three new haplotypes with restriction patterns of about 540, 420, 200, 150, 140 bp, about 540, 360, 280, 150, 140 bp, and about 580, 540, and 150, 140 bp have been identified along with the previously described haplotype X characterized by restriction products of 540, 420, 260, 150, and 140 bp in length.