Diagnostic Significance of Circulating EBV-miR-BHRF1-3 and EBV-miR-BART2-5p in Lymphoma A Case-Control Study

Epstein-Barr virus (EBV)-derived microRNAs (miRNAs) have been proposed as potential circulating biomarkers in EBV-associated malignancies. This study aimed to quantify the expression levels of four EBV-miRNAs (ebv-miR-BHRF1-2-5p, BHRF1-3, BART1-3p, and BART2-5p) in peripheral blood and to explore their possible diagnostic value in distinguishing lymphoma patients from healthy individuals.

This case control study included 50 newly diagnosed lymphoma patients (22 Hodgkin lymphoma, 28 Non-Hodgkin lymphoma) and 50 healthy controls recruited in 2025 at the Department of Internal Medicine (Hematology), Atatürk University. qRT-PCR analyses were performed at the Department of Medical Biology. EBV-miRNA expression levels were calculated using the ΔCt method with U6 snRNA as an internal control. Statistical analyses included FDR correction and ROC curve evaluation.

Ct values for U6 snRNA and all four EBV-miRNAs were significantly lower in lymphoma patients compared with controls (all p < 0.001, FDR < 0.001), indicating increased levels of circulating viral transcripts. ROC analyses showed that some miRNAs, particularly BHRF1-3 and BART1-3p, demonstrated notable discriminatory ability between patients and controls. No significant overall differences were observed between Hodgkin and Non-Hodgkin lymphoma; however, subtype analysis revealed significant variation in BHRF1-3 and BART2-5p expression (FDR = 0.005).

Circulating EBV-miRNAs were elevated in lymphoma patients, and several miRNAs showed promising discriminatory performance. However, this study is limited by its single-center design and the absence of external validation. Therefore, these findings should be considered preliminary, and further research in larger and independent cohorts is needed to confirm the diagnostic utility of EBV-derived miRNAs in lymphoma.