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Mosquitoes were collected in Colorado during 2006 and 2007 to examine spatial and seasonal patterns of risk for exposure to Culex vectors and West Nile virus. We used universal flavivirus primers to test pools of Culex mosquitoes for viral RNA. This led to the detection and subsequent isolation of two insect-specific flaviviruses: Culex flavivirus (CxFV), which was first described from Japan, and a novel insect flavivirus, designated Calbertado virus (CLBOV), which has also been detected in California and Canada. We recorded both viruses in Cx. tarsalis and Cx. pipiens from Colorado. Furthermore, quantitative reverse transcription polymerase chain reaction (RT-PCR) revealed the presence of CxFV RNA in Cx. pipiens eggs and larvae from a laboratory colony established in 2005 and naturally infected with CxFV, suggesting vertical transmission as a means of viral maintenance in natural Culex populations. Finally, we present phylogenetic analyses of the relationships between insect-specific flaviviruses and other selected flaviviruses.
Financial support: This research project was supported in part by contract N01-AI-25489 from the National Institutes of Allergy and Infectious Diseases.
Authors' addresses: Bethany G. Bolling, Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, Fort Collins, CO, E-mail: firstname.lastname@example.org. Lars Eisen, Chester G. Moore, and Carol D. Blair, Arthropod-Borne and Infectious Diseases Laboratory, Department of Microbiology, Immunology, and Pathology, Colorado State University, Fort Collins, CO, E-mails: email@example.com, firstname.lastname@example.org, and email@example.com.
Reprint requests: Bethany G. Bolling, Division of Vector-Borne Diseases, Centers for Disease Control and Prevention, 3150 Rampart Rd., Fort Collins, CO 80521, E-mail: firstname.lastname@example.org.