Multiplex qRT-PCR for the Detection of Western Equine Encephalomyelitis, St. Louis Encephalitis, and West Nile Viral RNA in Mosquito Pools (Diptera: Culicidae)

Journal of Medical Entomology
Aaron C BraultWilliam K Reisen

Abstract

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.

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Citations

Nov 19, 2015·Expert Review of Molecular Diagnostics·Robert DuncanMing Dong
Aug 6, 2016·Journal of Medical Entomology·Brittany M NelmsJamesina J Scott
Mar 13, 2016·Journal of Invertebrate Pathology·P GraystockW O H Hughes
Dec 12, 2018·Journal of Medical Entomology·Lucy L RobbRebekah C Kading
Feb 5, 2019·PLoS Neglected Tropical Diseases·Gabriella WorwaWilliam K Reisen
Sep 25, 2019·Journal of Medical Entomology·Laura D KramerA Marm Kilpatrick
Jun 11, 2020·PLoS Neglected Tropical Diseases·Daniele M SwetnamLark L Coffey
Nov 22, 2016·Emerging Infectious Diseases·Gregory S WhiteLark L Coffey
Aug 21, 2018·Emerging Infectious Diseases·Charsey Cole PorseVicki Kramer
Feb 26, 2019·Journal of Veterinary Diagnostic Investigation : Official Publication of the American Association of Veterinary Laboratory Diagnosticians, Inc·Hanna BristerNicola Pusterla
Jan 4, 2018·Journal of Zoo and Wildlife Medicine : Official Publication of the American Association of Zoo Veterinarians·Nicole CaptanianWilliam K Reisen
Nov 19, 2020·PLoS Neglected Tropical Diseases·Robert E SnyderVicki L Kramer
Aug 8, 2021·Animals : an Open Access Journal From MDPI·Alexandra Knox, Travis Beddoe

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