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

  • Lanciotti R
  • Kerst A
  • Nasci R
 et al. 
  • 4

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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.

Author-supplied keywords

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

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Authors

  • R S Lanciotti

  • A J Kerst

  • R S Nasci

  • M S Godsey

  • C J Mitchell

  • H M Savage

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