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

      Journal of Clinical Microbiology

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

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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.

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          Journal
          11060069
          87542

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