{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2024,7,26]],"date-time":"2024-07-26T19:54:31Z","timestamp":1722023671913},"reference-count":24,"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>Monitoring of Epstein\u2013Barr virus (EBV) infection and reactivation in immunocompromized patients (e.g., after organ or bone\u2010marrow transplantation) is based mainly on serological assays and detection of viral DNA. For further characterization of virus reactivation and monitoring of viral transcription we established real\u2010time RT\u2010PCR assays using TaqMan technology to sensitively quantify viral transcripts expressed at different times of the lytic cycle: for BZLF1, an immediate early transactivator initiating the transition from latency to lytic replication, for the DNA\u2010polymerase BALF5 and for the major viral glycoprotein gp350\/220 (BLLF1). RNA\u2010isolation was optimized to eliminate contaminating DNA. Preparations were shown to be virtually DNA\u2010free for up to 10<jats:sup>6<\/jats:sup> copies of RNA. With our PCR systems, it is possible to detect 10 copies of DNA or 100 copies of RNA per reaction as shown with serial dilutions of DNA\u2010plasmids or in vitro transcribed RNA, respectively. This corresponds to a detection limit of 8\u2009\u00d7\u200910<jats:sup>2<\/jats:sup> copies\/10<jats:sup>6<\/jats:sup> peripheral blood mononuclear cells (PBMCs). Evaluation of this system showed that even in healthy carriers borderline levels of BLLF1 mRNA were sometimes detectable. In patients with acute infectious mononucleosis (IM) viral transcripts were regularly found in varying concentrations. Extremely high levels of all three mRNA species could be seen in a patient after bone\u2010marrow transplantation monitored during an episode of lymphoproliferation which regressed during treatment with acyclovir and transfusion of donor T\u2010cells. This sensitive and reproducible method to detect and quantify different transcripts of EBV can be used to closely monitor reactivation of EBV, e.g., in immunocompromized patients. J. Med. Virol. 74:612\u2013618, 2004. \u00a9 2004 Wiley\u2010Liss, Inc.<\/jats:p>","DOI":"10.1002\/jmv.20220","type":"journal-article","created":{"date-parts":[[2004,10,13]],"date-time":"2004-10-13T17:38:29Z","timestamp":1097689109000},"page":"612-618","source":"Crossref","is-referenced-by-count":17,"title":["Quantitation of Epstein\u2013Barr virus mRNA using reverse transcription and real\u2010time PCR"],"prefix":"10.1002","volume":"74","author":[{"given":"Birgit","family":"Weinberger","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Annelie","family":"Plentz","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Klaus 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