title: Development of conventional and real-time reverse transcription polymerase chain reaction assays to detect Tembusu virus in Culex tarsalis mosquitoes.
abstract: Tembusu virus (TMUV) is an important emerging arthropod-borne virus that may cause encephalitis in humans and has been isolated in regions of southeast Asia, including Malaysia, Thailand, and China. Currently, detection and identification of TMUV are limited to research laboratories, because quantitative rapid diagnostic assays for the virus do not exist. We describe the development of sensitive and specific conventional and real-time quantitative reverse transcription polymerase chain reaction assays for detecting TMUV RNA in infected cell culture supernatant and Culex tarsalis mosquitoes. We used this assay to document the replication of TMUV in Cx. tarsalis, where titers increased 1,000-fold 5 days after inoculation. These assays resulted in the detection of virus-specific RNA in the presence of copurified mosquito nucleic acids. The use of these rapid diagnostic assays may have future applications for field pathogen surveillance and may assist in early detection, diagnosis, and control of the associated arthropod-borne pathogens.
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Lawrence N
authors.initials: LN
authors.lastname: Petz
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland michael.j.turell@us.army.mil.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Turell
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Susana
authors.initials: S
authors.lastname: Padilla
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Lewis S
authors.initials: LS
authors.lastname: Long
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Drew D
authors.initials: DD
authors.lastname: Reinbold-Wasson
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Darci R
authors.initials: DR
authors.lastname: Smith
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Monica L
authors.initials: ML
authors.lastname: O'Guinn
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: Vanessa R
authors.initials: VR
authors.lastname: Melanson
authors.affiliation: Virology Division, US Army Medical Research Institute of Infectious Diseases, Fort Detrick, Maryland; US Army Medical Research and Materiel Command, Fort Detrick, Maryland.
authors.forename: John S
authors.initials: JS
authors.lastname: Lee
mesh_terms: D000818:Animals; D001483:Base Sequence; D002522:Chlorocebus aethiops; D003465:Culex / Q000821:virology*; D017931:DNA Primers / Q000235:genetics; D005416:Flavivirus / Q000235:genetics / Q000302:isolation & purification*; D018177:Flavivirus Infections / Q000175:diagnosis* / Q000821:virology; D006801:Humans; D007303:Insect Vectors / Q000821:virology*; D008969:Molecular Sequence Data; D012367:RNA, Viral / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D013997:Time Factors; D014709:Vero Cells; D014779:Virus Replication
chemical_list: D017931:DNA Primers; D012367:RNA, Viral
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