The new Internal Transcribed Spacer 2 diagnostic tool clarifies the taxonomic position and geographic distribution of the North American malaria vector Anopheles punctipennis

Abstract Background The malaria mosquito Anopheles punctipennis , a widely distributed species in North America, is capable of transmitting human and actively involved the transmission ungulate parasite Plasmodium odocoilei . However, molecular diagnostic tools based on Internal Transcribed Spacer 2 (ITS2) ribosomal DNA are lacking for this species. former member maculipennis complex but its systematic position remains unclear. Methods In study, ITS2 sequences were obtained from 276 An. specimens collected eastern midwestern United States simple robust Restriction Fragment Length Polymorphism approach identification was developed. maximum-likelihood phylogenetic tree constructed available through study GenBank 20 Anopheles. Results analysis demonstrated consistent sequence length showed no indications intragenomic variation among samples ITS2, suggesting that represents single studied geographic locations. found urban, rural, forest settings, potential broad role pathogen transmission. Phylogeny comparison close relationship with other members Maculipennis group. Conclusions This developed vector clarified within