Multiplex qRT-PCR for the Detection of Western Equine Encephalomyelitis, St. Louis Encephalitis, and West Nile Viral RNA in Mosquito Pools (Diptera: Culicidae).
Following the introduction of West Nile virus into California during the summer of 2003, public health and vector control programs expanded surveillance efforts and were in need of diagnostics capable of rapid, sensitive, and specific detection of arbovirus infections of mosquitoes to inform decision support for intervention. Development of a multiplex TaqMan or real-time semiquantitative reverse transcription polymerase chain reaction (RT-PCR) assay in which three virus specific primer-probe sets were used in the same reaction is described herein for the detection of western equine encephalomyelitis, St. Louis encephalitis and West Nile viral RNA. Laboratory validation and field data from 10 transmission seasons are reported. The comparative sensitivity and specificity of this multiplex assay to singleplex RT-PCR as well as an antigen detection (rapid analyte measurement platform) and standard plaque assays indicate this assay to be rapid and useful in providing mosquito infection data to estimate outbreak risk.
Center for Vectorborne Diseases and Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616. Division of Vector-Borne Diseases, Centers for Diseases Control and Prevention, Fort Collins, CO 80512.
Aaron C
AC
Brault
Center for Vectorborne Diseases and Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616.
Ying
Y
Fang
Center for Vectorborne Diseases and Department of Pathology, Microbiology and Immunology, School of Veterinary Medicine, University of California, Davis, CA 95616. wkreisen@ucdavis.edu.
William K
WK
Reisen
D000818:Animals; D002140:California; D009033:Culicidae / Q000821:virology*; D004665:Encephalitis Virus, St. Louis / Q000302:isolation & purification*; D004667:Encephalitis Virus, Western Equine / Q000302:isolation & purification*; D004674:Encephalitis, St. Louis / Q000635:transmission / Q000821:virology; D020241:Encephalomyelitis, Western Equine / Q000635:transmission / Q000821:virology; D060885:Multiplex Polymerase Chain Reaction; D012367:RNA, Viral / Q000032:analysis*; D012680:Sensitivity and Specificity; D014901:West Nile Fever / Q000635:transmission / Q000821:virology; D014902:West Nile virus / Q000302:isolation & purification*
D012367:RNA, Viral
St. Louis encephalitis virus; West Nile virus; multiplex; surveillance; western equine encephalitis virus