Rapid detection and typing of dengue viruses from clinical samples by using reverse transcriptase-polymerase chain reaction

DOI Web Site 被引用文献28件
  • R S Lanciotti
    Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, Colorado 80522.
  • C H Calisher
    Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, Colorado 80522.
  • D J Gubler
    Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, Colorado 80522.
  • G J Chang
    Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, Colorado 80522.
  • A V Vorndam
    Division of Vector-Borne Infectious Diseases, Centers for Disease Control, Fort Collins, Colorado 80522.

書誌事項

公開日
1992-03
権利情報
  • https://journals.asm.org/non-commercial-tdm-license
DOI
  • 10.1128/jcm.30.3.545-551.1992
公開者
American Society for Microbiology

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説明

<jats:p>We report on the development and application of a rapid assay for detecting and typing dengue viruses. Oligonucleotide consensus primers were designed to anneal to any of the four dengue virus types and amplify a 511-bp product in a reverse transcriptase-polymerase chain reaction (PCR). First, we produced a cDNA copy of a portion of the viral genome in a reverse transcriptase reaction in the presence of primer D2 and then carried out a standard PCR (35 cycles of heat denaturation, annealing, and primer extension) with the addition of primer D1. The resulting double-stranded DNA product of the RT-PCR was typed by two methods: dot blot hybridization of the 511-bp amplified product to dengue virus type-specific probes or a second round of PCR amplification (nested PCR) with type-specific primers, yielding DNA products the unique sizes of which were diagnostic for each dengue virus serotype. The accumulated data demonstrated that dengue viruses can be accurately detected and typed from viremic human serum samples.</jats:p>

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