Weinberger, Birgit and Plentz, Annelie and Weinberger, Klaus M. and Hahn, Joachim and Holler, Ernst and Jilg, Wolfgang (2004) Quantitation of Epstein-Barr virus MRNA using reverse transcription and real-time PCR. JOURNAL OF MEDICAL VIROLOGY, 74 (4). pp. 612-618. ISSN 0146-6615,
Full text not available from this repository. (Request a copy)Abstract
Monitoring of Epstein-Barr virus (EBV) infection and reactivation in immunocompromized patients (e.g., after organ or bone-marrow 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-time RTPCR 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-polymerase BALF5 and for the major viral glycoprotein gp350/220 (BLLF1). RNA-isolation was optimized to eliminate contaminating DNA. Preparations were shown to be virtually DNA-free for up to 10(6) 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-plasmids or in vitro transcribed RNA, respectively. This corresponds to a detection limit of 8 x 10(2) copies/10(6) 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-marrow transplantation monitored during an episode of lymphoproliferation which regressed during treatment with acyclovir and transfusion of donor T-cells. 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.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | BONE-MARROW-TRANSPLANTATION; POSTTRANSPLANT LYMPHOPROLIFERATIVE DISORDER; GENE-EXPRESSION; ENVELOPE GLYCOPROTEINS; PERIPHERAL-BLOOD; CARRIER STATE; RISK-FACTORS; RECIPIENTS; DNA; DISEASE; lytic cycle; viral reactivation; TaqMan-PCR; lymphoproliferation |
| Subjects: | 600 Technology > 610 Medical sciences Medicine |
| Divisions: | Medicine > Lehrstuhl für Innere Medizin III (Hämatologie und Internistische Onkologie) Medicine > Lehrstuhl für Medizinische Mikrobiologie und Hygiene |
| Depositing User: | Dr. Gernot Deinzer |
| Date Deposited: | 21 Jun 2021 10:54 |
| Last Modified: | 21 Jun 2021 10:54 |
| URI: | https://pred.uni-regensburg.de/id/eprint/36912 |
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