Detection of flaviviruses by reverse-transcriptase polymerase chain reaction

Z. A. Eldadah, D. M. Asher, M. S. Godec, K. L. Pomeroy, L. G. Goldfarb, S. M. Feinstone, H. Levitan, C. J. Gibbs, D. C. Gajdusek

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58 Scopus citations


RNA sequences of five flaviviruses were detected by a modified polymerase chain reaction (PCR) that incorporated a reverse transcriptase and RNase inhibitor. Oligonucleotide primer pairs were synthesized to amplify sequences from St. Louis encephalitis (SLE), Japanese encephalitis (JBE), yellow fever (YF), dengue 2 (DEN-2), and dengue 4 (DEN-4) viruses. The amplified products were visualized as bands of appropriate size on ethidium bromide-stained agarose gels. The identity of these products was confirmed by restriction endonuclease cleavage to generate fragments of predicted lengths. The reverse-transcriptase PCR (RT-PCR) successfully amplified flavivirus sequences from cell cultures, frozen brain tissue, and formalin-fixed, paraffin-embedded brain tissue. The reactions were highly specific, and the method compared favorably to two conventional assays of viral infectivity. RT-PCR followed by PCR with nesting primers (N-PRC) was 1,000-fold more sensitive in detecting virus than classical infectivity titration by intracerebral inoculation of suckling mice and nearly 1,000-fold more sensitive than amplification of virus in cell culture followed by inoculation of mice.

Original languageEnglish (US)
Pages (from-to)260-267
Number of pages8
JournalJournal of Medical Virology
Issue number4
StatePublished - 1991
Externally publishedYes


  • Dengue
  • Japanese encephalitis
  • Nesting primers
  • Reverse transcriptase
  • RNA
  • St. Louis encephalitis
  • Yellow fever

ASJC Scopus subject areas

  • Virology


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