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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)

Detection of flaviviruses by reverse-transcriptase polymerase chain reaction.

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-PCR) 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.[1]


  1. Detection of flaviviruses by reverse-transcriptase polymerase chain reaction. Eldadah, Z.A., Asher, D.M., Godec, M.S., Pomeroy, K.L., Goldfarb, L.G., Feinstone, S.M., Levitan, H., Gibbs, C.J., Gajdusek, D.C. J. Med. Virol. (1991) [Pubmed]
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