A Feasible Flowchart to Screen EBV-Positive Diffuse Large B Cell Lymphoma of the elderly: A Molecular Approach

Background: Epstein Barr Virus-positive diffuse large B cell lymphoma of the elderly (DLBCL-EBV + - e ), initially described in 2003, is a provisional entity in the 4 th edition of the World Health Organization (WHO) Classification of tumors of hematopoietic and lymphoid tissues and is defined as an EBV + monoclonal large B cell proliferation that occurs in patients >50 years of age and in whom there is no known immunodeficiency or history of lymphoma. It is associated with a poor prognosis but could be treated with the association of novel therapeutic approaches. EBER in situ hybridization (EBER-ISH)  is the gold standard assay for defining a tumor as EBV-related, however there are no uniform criteria for the percentage of EBV-positive cells in DLBCL-EBV + - e . Methods: We applied to Formalin Fixed Paraffin Embedded (FFPE) tissue of 43 novel DLBCL over 50 years Q-PCR for the detection of viral DNA in concert with EBER-ISH. Result: Only one case was defined as true DLBCL-EBV+- e being Q-PCR positive  with a value of log EBV DNA/10 5 cells ≥ 6 and EBER-ISH positive. Conclusion: In the current study we applied to FFPE tissue of DLBCL-EBV + - e a reliable and feasible Q-PCR/EBER-ISH combined protocol for the screening of EBV status. Our protocol could help to better identify the DLBCL-EBV + - e patients eligible for novel therapeutic approaches.