Validation of Roche LightCycler Epstein-Barr virus quantification reagents in a clinical laboratory setting

Margaret L. Gulley, Hongxin Fan, Sandra H. Elmore

Research output: Contribution to journalArticlepeer-review

18 Scopus citations

Abstract

Epstein-Barr virus (EBV) is associated with a wide range of benign and malignant diseases, including infectious mononucleosis, lymphoma, posttransplant lymphoproliferative disorder, and nasopharyngeal carcinoma. Measurement of EBV viral load in plasma is increasingly used for rapid assessment of disease status. We evaluated the performance characteristics of an EBV polymerase chain reaction assay that uses commercial reagents and instruments from Roche Diagnostics (Indianapolis, IN). DNA was extracted from plasma using a MagNaPure instrument, and viral load was measured by real-time polymerase chain reaction on a LightCycler. Analyte-specific re-agents included primers and hybridization probes targeting the EBV LMP2 gene and a spiked control sequence. Accuracy and reproducibility were established using DNA from three cell lines. The assay was sensitive to approximately 750 copies of EBV DNA per milliliter of plasma and was linear across at least four orders of magnitude. The assay detected EBV DNA in three of five samples from nasopharyngeal carcinoma patients, seven of nine infectious mononucleosis samples, and 34/34 samples from immunosuppressed patients with clinically significant EBV-related disease, whereas EBV DNA was undetectable in plasma from 21 individuals without EBV-related disease. In conclusion, this LightCycler EBV assay is rapid, sensitive, and linear for quantifying EBV viral load. The assay appears to be usefull for measuring clinically significant EBV levels in immunodeficient patients.

Original languageEnglish (US)
Pages (from-to)589-597
Number of pages9
JournalJournal of Molecular Diagnostics
Volume8
Issue number5
DOIs
StatePublished - Nov 2006

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Medicine

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