A real-time TaqMan polymerase chain reaction for the identification of Culex vectors of West Nile and Saint Louis encephalitis viruses in North America.
In North America, West Nile and St. Louis encephalitis viruses have been detected in a wide range of vector species, but the majority of isolations continue to be from pools of mixed mosquitoes in the Culex subgenus Culex. Unfortunately, the morphologic identification of these important disease vectors is often difficult, particularly in regions of sympatry. We developed a sensitive real-time TaqMan polymerase chain reaction assay that allows reliable identification of Culex mosquitoes including Culex pipiens pipiens, Cx. p. quinquefasciatus, Cx. restuans, Cx. salinarius, Cx. nigripalpus, and Cx. tarsalis. Primers and fluorogenic probes specific to each species were designed based on sequences of the acetylcholinesterase gene (Ace2). Both immature and adult mosquitoes were successfully identified as individuals and as mixed species pools. This identification technique provides the basis for a rapid, sensitive, and high-throughput method for expounding the species-specific contribution of vectors to various phases of arbovirus transmission.
Sanogo
Yibayiri O
YO
Illinois Natural History Survey, Champaign, Illinois 61820, USA. sanogo@uiuc.edu
Kim
Chang-Hyun
CH
Lampman
Richard
R
Novak
Robert J
RJ
D000818:Animals; D003465:Culex / Q000235:genetics* / Q000821:virology; D017931:DNA Primers; D004665:Encephalitis Virus, St. Louis / Q000302:isolation & purification*; D004674:Encephalitis, St. Louis / Q000635:transmission*; D006801:Humans; D007303:Insect Vectors / Q000235:genetics* / Q000821:virology; D009656:North America; D016133:Polymerase Chain Reaction / Q000379:methods*; D011237:Predictive Value of Tests; D012680:Sensitivity and Specificity; D014901:West Nile Fever / Q000635:transmission*; D014902:West Nile virus / Q000302:isolation & purification*
D017931:DNA Primers