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Three sets of two primers and one internal dual\u2010labeled probe for each primer set that target distinct conserved regions of the rabies virus N gene were designed and evaluated. Using a collection of 203 isolates representative of the world\u2010wide diversity of rabies virus, all three primers\/probe sets were shown to detect a wide range of rabies virus strains with very few detection failures; the RABVD1 set in particular was the most broadly reactive. These qRT\u2010PCR assays were shown to be quantitative over a wide range of viral titer and were 100\u20131,000 times more sensitive than nested RT\u2010PCR; however, both the standard and real\u2010time PCR methods yielded concordant results when used to test a collection of archived human suspect samples. The qRT\u2010PCR assay was employed to monitor virus load in the saliva of a rabies virus\u2010infected patient undergoing the Milwaukee treatment protocol. However in this case it would appear that reduction of the viral load in the patient's saliva over time did not appear to correlate well with clearance of viral components from the brain. J. Med. Virol. 81:1484\u20131497, 2009. \u00a9 2009 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/jmv.21547","type":"journal-article","created":{"date-parts":[[2009,6,23]],"date-time":"2009-06-23T12:22:45Z","timestamp":1245759765000},"page":"1484-1497","source":"Crossref","is-referenced-by-count":79,"title":["Development of real\u2010time reverse transcriptase polymerase chain reaction methods for human rabies diagnosis"],"prefix":"10.1002","volume":"81","author":[{"given":"Susan A.","family":"Nadin\u2010Davis","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mary","family":"Sheen","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Alexander 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