title: Combining reverse-transcription multiplex PCR and microfluidic electrophoresis to simultaneously detect seven mosquito-transmitted zoonotic encephalomyelitis viruses.
abstract: Several mosquito-transmitted viruses are causative agents for zoonotic encephalomyelitis. Rapid identification of these viruses in mosquito populations is an effective method for surveying these diseases. To detect multiple mosquito-transmitted viral agents, including West Nile virus, Saint Louis encephalitis virus, Venezuelan equine encephalomyelitis virus, Western equine encephalomyelitis virus, Eastern equine encephalomyelitis virus, Highlands J virus and Japanese encephalitis virus, an assay using multiplex reverse-transcription PCR combined with microfluidic electrophoresis was developed and evaluated. Tailed nested primers were used in the assay to amplify specific viral genomic segments, and products with specific length were further analyzed by using a microfluidic electrophoresis chip. The assay exhibited good specificity and analytical sensitivity (10(2) copies/µL). This technology can be helpful in the quarantine and surveillance of exotic encephalomyelitis viruses which are transmitted by mosquitoes.
authors.affiliation: School of Life Sciences, Chongqing University, Chongqing 400030, China; Technical Center of Chongqing Entry Inspection and Quarantine Bureau, Engineering Research Center for Import and Export Food Safety, Chongqing 400020, China.
authors.forename: Yu
authors.initials: Y
authors.lastname: Wang
authors.affiliation: Diagnostic Virology Laboratory, National Veterinary Services Laboratories, STAS/VS/APHIS/USDA, Ames, IA 50010, USA.
authors.forename: Eileen N
authors.initials: EN
authors.lastname: Ostlund
authors.affiliation: Technical Center of Chongqing Entry Inspection and Quarantine Bureau, Engineering Research Center for Import and Export Food Safety, Chongqing 400020, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Jun
authors.affiliation: Technical Center of Chongqing Entry Inspection and Quarantine Bureau, Engineering Research Center for Import and Export Food Safety, Chongqing 400020, China.
authors.forename: Fu-Ping
authors.initials: FP
authors.lastname: Nie
authors.affiliation: School of Life Sciences, Chongqing University, Chongqing 400030, China; Technical Center of Chongqing Entry Inspection and Quarantine Bureau, Engineering Research Center for Import and Export Food Safety, Chongqing 400020, China.
authors.forename: Ying-Guo
authors.initials: YG
authors.lastname: Li
authors.affiliation: Diagnostic Virology Laboratory, National Veterinary Services Laboratories, STAS/VS/APHIS/USDA, Ames, IA 50010, USA.
authors.forename: Donna J
authors.initials: DJ
authors.lastname: Johnson
authors.affiliation: School of Life Sciences, Chongqing University, Chongqing 400030, China.
authors.forename: Rui
authors.initials: R
authors.lastname: Lin
authors.affiliation: School of Life Sciences, Chongqing University, Chongqing 400030, China. Electronic address: zhengguoli@cqu.edu.cn.
authors.forename: Zheng-Guo
authors.initials: ZG
authors.lastname: Li
mesh_terms: D000818:Animals; D009033:Culicidae / Q000821:virology*; D046209:Electrophoresis, Microchip / Q000662:veterinary*; D004668:Encephalitis Viruses / Q000302:isolation & purification*; D062665:Epidemiological Monitoring / Q000662:veterinary*; D060885:Multiplex Polymerase Chain Reaction / Q000662:veterinary*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000662:veterinary*; D017423:Sequence Analysis, RNA / Q000662:veterinary
keywords: Encephalomyelitis viruses; Microfluidic; Mosquito-transmitted viruses; Multiplex PCR