Detection of flaviviruses by reverse-transcriptase polymerase chain reaction.

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