Evaluation of a Quantitative Dual-Target EBV DNA Test on a Fully Automated Molecular Testing System

Area of Science:

• Virology
• Molecular Diagnostics
• Transplant Medicine

Background:

• Epstein-Barr virus (EBV) deoxyribonucleic acid (DNA) measurement is crucial for managing EBV-associated complications in transplant recipients.
• Accurate and sensitive EBV DNA quantification is essential for patient care.

Purpose of the Study:

• To evaluate the performance of the new Roche cobas EBV real-time PCR assay.
• To assess the assay's accuracy, sensitivity, specificity, and reproducibility for detecting EBV DNAemia.

Main Methods:

• Utilized EDTA-plasma dilution panels and spiked clinical samples with WHO EBV international standard or high-titer EBV lambda stock.
• Assessed correlation with the Abbott Realtime EBV assay in clinical specimens across two independent laboratories.
• Performed in silico analysis of the dual-target (EBNA1 and BMRF2) assay for EBV diversity inclusivity.

Main Results:

• The assay demonstrated high inclusivity for EBV diversity, with limits of detection (LoD) of 16.6 IU/mL (GT1) and 18.8 IU/mL (GT2).
• Linear ranges were wide, and accuracy was confirmed (mean difference ≤ ±0.18 log10).
• High correlation (R² = 0.96) was observed with a comparator assay in clinical samples, with a 0.1 mean log10 titer difference.

Conclusions:

• The Roche cobas EBV test is an accurate, sensitive, specific, and reproducible assay for detecting EBV DNAemia in plasma.
• Automation and calibration to international standards can improve harmonization of quantitative EBV DNA testing across laboratories.