Hodgkin lymphoma (HL) is commonly associated with latent Epstein-Barr virus (EBV) infection. The aim of this study is a molecular analysis of the EBV status in both involved lymph node biopsies and plasma samples of patients with HL. Plasma and lymph node biopsy samples obtained from 15 pediatric and 10 adult HL patients were examined for EBV DNA using conventional polymerase chain reaction (PCR). The control group consisted of 30 healthy pediatrics and adults. In addition, immunoglobulin G (IgG) anti-EBV nuclear antigen (EBNA)-1 antibody was determined in sera of patients and controls using enzyme-linked immunosorbent assay (ELISA). IgG Anti EBNA-1 antibody was detected in 21 (84%) and 8 (26%) of patients and controls, respectively (P < .05). EBV DNA was detected in 12 (48%) and 1 (3%) plasma samples of patient and control cases, respectively. Significant difference was observed in plasma DNA detection between patients and controls (P < .05). Comparison of EBV DNA detection in plasma and biopsy samples between children and adult patients was only significant for plasma samples (P = .025). Significant correlation was observed in positive detection of EBV DNA between plasma and biopsy samples of the same individual (P < .001, r = .923). Frequency of EBV DNA in plasma and biopsy samples obtained from mixed-cellularity subgroup was higher than the nodular sclerosis; however, no significant difference was observed between these 2 subgroups. EBV detection in plasma of childhood Hodgkin lymphoma in a population with EBV seroconversion might be of value as a biomarker for EBV-associated Hodgkin lymphoma.