Background: Epstein–Barr Virus (EBV) is one of the human gamma herpes viruses that promotes subclinical and latent infections in healthy B lymphocytes. This research objects to find out the value of expression of BHRF1-1 microRNA of EBV in chronic B- lymphocytic leukemia (B-CLL) patients. Methods: This retrospective research involved 40 subjects diagnosed with B-CLL and positive for Epstein Barr Virus Nuclear Antigen-1 IgG (EBNA-1 IgG) and 40 healthy volunteers serving as the control group. Subjects were allocated equally into two groups: group I: 40 patients with B-CLL  and group II: 40 healthy volunteers matched for age and sex with patients serving as a control group. Results: There was a significant increase in BHRF1-1 expression in CLL cases than control group (P value <0.001). Overall survival was significantly higher in patients with low BHRF1-1 expression (<3.63) than the patients with overexpression of BHRF1-1 in B-cells that was related to shorter OS BHRF1-1 expression (>3.63) (p value 0.004). Lymphocyte count (≥50 x109/L), CD38 Expression, Rai Staging (III/ IV), FISH (17p deletion vs del 13q14, trisomy 12 and del 11q23) and relative expression of BHRF1-1(high) were the significant indicators. Conclusions: EBV-encoded miRNAs are highly found in EBV-associated tumors as BHRF1-1 MicroRNA marker of EBV.