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Oligonucleotide primer pairs were synthesized to amplify sequences from St. Louis encephalitis (SLE), Japanese encephalitis (JBE), yellow fever (YF), dengue 2 (DEN\u20102), and dengue 4 (DEN\u20104) viruses. The amplified products were visualized as bands of appropriate size on ethidium bromide\u2010stained agarose gels. The identity of these products was confirmed by restriction endonuclease cleavage to generate fragments of predicted lengths. The reverse transcriptase PCR (RT\u2010PCR) successfully amplified flavivirus sequences from cell cultures, frozen brain tissue, and formalin\u2010fixed, paraffin\u2010embedded brain tissue. The reactions were highly specific, and the method compared favorably to two conventional assays of viral infectivity. RT\u2010PCR followed by PCR with nesting primers (N\u2010PCR) was 1,000\u2010fold more sensitive in detecting virus than classical infectivity titration by intracerebral inoculation of suckling mice and nearly 1,000\u2010fold more sensitive than amplification of virus in cell culture followed by inoculation of mice.<\/jats:p>","DOI":"10.1002\/jmv.1890330410","type":"journal-article","created":{"date-parts":[[2007,3,2]],"date-time":"2007-03-02T19:17:47Z","timestamp":1172863067000},"page":"260-267","source":"Crossref","is-referenced-by-count":59,"title":["Detection of flaviviruses by reverse\u2010transcriptase polymerase chain reaction"],"prefix":"10.1002","volume":"33","author":[{"given":"Zayd A.","family":"Eldadah","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"David M.","family":"Asher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mark S.","family":"Godec","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Kitty L.","family":"Pomeroy","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Lev G.","family":"Goldfarb","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stephen M.","family":"Feinstone","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Herbert","family":"Levitan","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"suffix":"Jr.","given":"C. 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