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Am. J. Trop. Med. Hyg., 77(1), 2007, pp. 58-66
Copyright © 2007 by The American Society of Tropical Medicine and Hygiene

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A Real-Time TaqMan Polymerase Chain Reaction for the Identification of Culex Vectors of West Nile and Saint Louis Encephalitis Viruses in North America

Yibayiri O. Sanogo*, Chang-Hyun Kim, Richard Lampman, AND Robert J. Novak
Illinois Natural History Survey, Champaign, Illinois

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.

Received October 31, 2006. Accepted for publication March 25, 2007.

Acknowledgments: The authors thank the team of the Champaign-Urbana Encephalitis Prevention Program for help with mosquito rearing and identification. The authors thank Dr. J. Anderson (The Connecticut Agricultural Experiment Station, New Haven, CT), Dr. R. Xue and Whitney Qualls (Anastasia Mosquito Control District, FL), and H. Lothrop (Coachella Valley Mosquito and Vector Control District, CA) and C. Palmisano (Saint Tammany Parish Mosquito Abatement District, LA) for providing the mosquito strains.

Financial support: This study was supported by a grant from the Illinois Waste Tire Fund and USDA/CREES Grant 2005-34523-15639 to R. J. Novak.

* Address correspondence to Yibayiri O. Sanogo, Illinois Natural History Survey, 1816 South Oak Street, Champaign, IL 61820. E-mail: sanogo{at}uiuc.edu

Authors’ address: Yibayiri O. Sanogo, Chang-Hyun Kim, Richard Lampman, and Robert J. Novak, Illinois Natural History Survey, 1816 South Oak Street, Champaign, IL 61820.






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Copyright © 2007 by the American Society of Tropical Medicine and Hygiene.