Application of Polymerase Chain Reaction-restriction Fragment Length Polymorphism Techniques in Differential Identification of Close-breeding Anopheline Mosquitoes from Anopheles hyrcanus Complex

Objective To identify species of anopheles vectors captured in varied areas of Hubei province with polymerase chain reaction-restriction fragment length polymorphism(PCR-RFLP) techniques.Methods Anopheline mosquitoes(Anopheles hyrcanus complex) captured at the fields of Hubei province were identified with traditional morphological classification and newly-established genetic methods(PCR-RFLP) based on the sequences of their ribosomal DNA in the ITS_2 region.DNA sequence in Anopheles anthropophagus,Anopheles sinensis and Anopheles yatsushiroensisi were analyzed and compared. Results(A total) of 181 anopheline mosquitoes were captured and identified,including 176 identified as Anopheles sinensis and five as Anopheles anthropophagus according to their morphological features,and 172 identified as Anopheles sinensis,four as Anopheles anthropophagus and one as Anopheles yatsushiroensisi by PCR-RFLP.Conclusion Close-breeding anopheles,such as Anopheles sinensis,Anopheles yatsushiroensisi and Anopheles anthropophagus in Anopheles hyrcanus complex could be accurately identified by PCR-RFLP,which was suitable for the investigation and surveillance for malaria-vector mosquitoes in the areas with anopheles complex.