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Development of conventional and real-time reverse transcription polymerase chain reaction assays to detect Tembusu virus in Culex tarsalis mosquitoes.

Authors
  • Petz, Lawrence N
  • Turell, Michael J
  • Padilla, Susana
  • Long, Lewis S
  • Reinbold-Wasson, Drew D
  • Smith, Darci R
  • O'Guinn, Monica L
  • Melanson, Vanessa R
  • Lee, John S
Type
Published Article
Journal
American Journal of Tropical Medicine and Hygiene
Publisher
American Society of Tropical Medicine and Hygiene
Publication Date
Oct 01, 2014
Volume
91
Issue
4
Pages
666–671
Identifiers
DOI: 10.4269/ajtmh.13-0218
PMID: 25114013
Source
Medline
License
Unknown

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.

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