BRENGEL-PESCE K, MORAND P, BARGUES G, BUISSON M, SEIGNEURIN JM; Interscience Conference on Antimicrobial Agents and Chemotherapy.

Abstr Intersci Conf Antimicrob Agents Chemother Intersci Conf Antimicrob Agents Chemother. 2000 Sep 17-20; 40: 276.

J. Fourier Univ., Grenoble, France

BACKGROUND The measurement of EBV load in blood is thought to be useful to diagnose and monitor EBV-associated diseases particularly EBV-associated post-transplantation lymphoproliferative disorders or in some EBV-associated AIDS-lymphoma. This study presents the assessment of EBV load in peripheral blood mononuclear cells (PBMC) or sera from healthy or immunocompromised subjects (HIV or transplanted patients) with or without EBV-associated diseases with the real-time quantitative LightCycler PCR (LC-PCR) technology. METHODS LC-PCR was developed to amplify a 169bp fragment of EBV thymidine kinase gene from DNA extracted from PBMC and sera. Probes using Fluorescence Resonance Energy Transfer principle assured the specificity. RESULTS LC-PCR could detect 2 copies of EBV genome. The specificity was confirmed from EBV-negative cell lines, other human herpesviruses and EBV-seronegative individuals. A good reproducibility inter-assay of LC-PCR was obtained from 42 samples (r2=0.9664). LC-PCR results were compared with a routinely used ELISA-PCR assay from 150 samples and a good correlation was found (r2=0.8124). The majority of immunocompetent-EBV seropositive-healthy patients (24 out of 34, 70%) were negative for EBV DNA in PBMC, or had very low EBV DNA levels (mean of 11 copy/microg DNA ). None had EBV detected in serum. In contrast, the majority of patients with infectious mononucleosis had EBV DNA in serum (9 out of 12, 75%, mean of 1169 EBV copy/ml) and 80% immunocompromised patients without EBV associated disease were LC-PCR positive with a higher EBV DNA level in PBMC (mean of 469 copy) than healthy patients. Furthermore, the retrospective follow up of 5 transplanted patients with EBV-associated lymphoproliferative disease showed a peak of EBV viral load occurring in PBMC and/or serum before the appearance of the clinical disease and a decrease of this load under specific treatment. CONCLUSION This study demonstrates the practicability and the accuracy of the real-time quantitative LightCycler-PCR strategy for the monitoring of EBV viral load in a clinical setting.KEYWORDS: Epstein-Barr virus; Real-time quantitative; PCR

Publication Types:

• Meeting Abstracts

Keywords:

• Enzyme-Linked Immunosorbent Assay
• Herpesvirus 4, Human
• Humans
• Infectious Mononucleosis
• Lymphoproliferative Disorders
• Meditation
• Polymerase Chain Reaction
• Sensitivity and Specificity
• Time
• Viral Load
• transplantation

Other ID:

• GWAIDS0010434

UI: 102247932

From Meeting Abstracts