Two widespread human herpesviruses, the Epstein-Barr virus (EBV) and the Human Herpesvirus 6 (HHV-6), have been frequently associated with Hodgkin's Disease (HD) and, recently, it has been observed an HHV-6 transactivation effect on EBV replicative cycle. We studied the presence and the possible association between EBV and HHV-6 in childhood HD cases, nodular sclerosis subtype. We analyzed formalin-fixed and paraffin-embedded lymph nodes from 15 cases by PCR for HHV-6 genome, and by PCR and in situ hybridization (ISH) for EBV genome. One out 15 samples resulted positive for HHV-6 DNA PCR, while 5 resulted positive for EBV DNA PCR. Only one sample positive for HHV-6 resulted positive for both HHV-6 and EBV genome. All samples were negative in ISH. At the moment, it is not clear the exact role of EBV and HHV-6 in the lymphomagenesis, neither it is possible to establish the rate of their interaction; our data show that it does not exist in vivo an evidence of their association.