การพัฒนาเครื่องหมาย allele-specific PCR เพื่อจ าแนกชนิดยุงก้นปล่องกลุ่มซับซ้อน Anopheles dirus Development of Allele-specific PCR for Species Identification of the Anopheles dirus complex

Anopheles dirus complex is widely distributed as primary malaria vectors in Thailand and Southeast Asia region. Morphological identification is very problematic for the Dirus complex species because of their similar morphologies among the sibling species, which usually lead to species misidentification. In this research, allele-specific PCR was developed to identify the three species within Anopheles dirus complex group in Thailand. The allele-specific primers were designed based on the nucleotide polymorphisms in mitochondrial cytochrome oxidase subunit I sequences. The results showed that these primers clearly amplified Anopheles cracens, An. scanloni and An. nemophilous for different PCR products size of 236, 338 and 527 base pair, respectively. This preliminary result suggested that the allele-specific PCR could be correctly and effectively identified member of the Dirus species complex.