Rapid detection of west nile virus from human clinical specimens, field-collected mosquitoes, and avian samples by a TaqMan reverse transcriptase-PCR assay

J Clin Microbiol. 2000 Nov;38(11):4066-71. doi: 10.1128/JCM.38.11.4066-4071.2000.

Abstract

The authors report on the development and application of a rapid TaqMan assay for the detection of West Nile (WN) virus in a variety of human clinical specimens and field-collected specimens. Oligonucleotide primers and FAM- and TAMRA-labeled WN virus-specific probes were designed by using the nucleotide sequence of the New York 1999 WN virus isolate. The TaqMan assay was compared to a traditional reverse transcriptase (RT)-PCR assay and to virus isolation in Vero cells with a large number ( approximately 500) of specimens obtained from humans (serum, cerebrospinal fluid, and brain tissue), field-collected mosquitoes, and avian tissue samples. The TaqMan assay was specific for WN virus and demonstrated a greater sensitivity than the traditional RT-PCR method and correctly identified WN virus in 100% of the culture-positive mosquito pools and 98% of the culture-positive avian tissue samples. The assay should be of utility in the diagnostic laboratory to complement existing human diagnostic testing and as a tool to conduct WN virus surveillance in the United States.

Publication types

  • Evaluation Study

MeSH terms

  • Animals
  • Bird Diseases / diagnosis*
  • Bird Diseases / virology
  • Birds / virology
  • Brain / virology
  • Chlorocebus aethiops
  • Culicidae / virology*
  • Humans
  • RNA, Viral / blood
  • RNA, Viral / cerebrospinal fluid
  • Reverse Transcriptase Polymerase Chain Reaction*
  • Sensitivity and Specificity
  • Taq Polymerase / metabolism*
  • Vero Cells
  • Virus Cultivation
  • West Nile Fever / diagnosis*
  • West Nile Fever / veterinary
  • West Nile Fever / virology
  • West Nile virus / genetics
  • West Nile virus / isolation & purification*

Substances

  • RNA, Viral
  • Taq Polymerase