{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,21]],"date-time":"2026-07-21T21:54:49Z","timestamp":1784670889948,"version":"3.55.0"},"reference-count":46,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2004,10,13]],"date-time":"2004-10-13T00:00:00Z","timestamp":1097625600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Medical Virology"],"published-print":{"date-parts":[[2004,12]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:p>A rapid, sensitive, and accurate laboratory diagnostic test is needed for distinguishing Japanese encephalitis virus (JEV) from other diseases featuring similar clinical symptoms and also for preventing potential outbreaks. In this study, a TaqMan reverse transcription (RT)\u2010polymerase chain reaction (PCR) assay was developed for rapid detection and quantification of the viral RNA of various JEV strains. A consensus JEV NS3 region was chosen to design the primers and the TaqMan probe. The JEV TaqMan assay used the EZ\u2010r<jats:italic>TtH<\/jats:italic> RT\u2010PCR system featuring advantages such as a one\u2010step, high\u2010temperature RT reaction modality and preventing carry\u2010over contamination. The sensitivity of the JEV TaqMan assay for detecting in vitro\u2010transcribed JEV NS3 RNA was estimated to be one to five copies of RNA per reaction. For cultured JE virions, less than 40 plaque forming unit (PFU)\/ml of virus load (corresponding to 0.07 PFU\/test) could be detected. In addition, the JEV TaqMan assay could detect all seven strains of JEV tested, but provided negative results for nine other flaviviruses and encephalitis viruses tested. The JEV TaqMan assay demonstrated greater sensitivity and specificity than traditional RT\u2010PCR methods as has been previously reported. The application of the JEV TaqMan assay herein has been shown to the sensitive detection of the JEV from both mosquito pools and also JEV\u2010spiking human blood. The assay should be of use in diagnostic laboratory conduct and could be used to replace or complement time\u2010consuming viral\u2010culture methods, thus achieving more rapid, sensitive, and highly specific identification of JEV infection. J. Med. Virol. 74:589\u2013596, 2004. \u00a9 2004 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/jmv.20218","type":"journal-article","created":{"date-parts":[[2004,10,13]],"date-time":"2004-10-13T17:37:59Z","timestamp":1097689079000},"page":"589-596","source":"Crossref","is-referenced-by-count":35,"title":["Sensitive and specific detection of strains of Japanese encephalitis virus using a one\u2010step TaqMan RT\u2010PCR 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