A multiplex PCR-based molecular identification of five morphologically related, medically important subgenus Stegomyia mosquitoes from the genus Aedes (Diptera: Culicidae) found in the Ryukyu Archipelago, Japan.
Internal transcribed spacer regions of ribosomal DNA were sequenced, and new species-specific primers were designed to simplify the molecular identification of five morphologically related subgenus Stegomyia mosquito species--Aedes aegypti, Ae. albopictus, Ae. riversi, Ae. flavopictus, and Ae. daitensis--found in the Ryukyu Archipelago, Japan. Each newly designed primer was able to amplify a species-specific fragment with a different size. Conditions for multiplex PCR were optimized to identify all five species in a single PCR. This method is a convenient tool for entomological field surveys, particularly in arbovirus endemic/epidemic areas where some of these species coexist.
Higa
Yukiko
Y
Department of Vector Ecology and Environment, Institute of Tropical Medicicine, Nagasaki University, Nagasaki 852-8523, Japan. yukko@nagasaki-u.ac.jp
Toma
Takako
T
Tsuda
Yoshio
Y
Miyagi
Ichiro
I
D000330:Aedes / Q000235:genetics*; D000818:Animals; D017931:DNA Primers; D021903:DNA, Ribosomal Spacer / Q000235:genetics*; D004587:Electrophoresis, Agar Gel; D005260:Female; D005820:Genetic Testing / Q000379:methods; D007303:Insect Vectors / Q000235:genetics*; D007564:Japan; D016133:Polymerase Chain Reaction / Q000379:methods; D013045:Species Specificity
D017931:DNA Primers; D021903:DNA, Ribosomal Spacer