There are a number of reports regarding natural infection of Dermacentor reticulatus ticks with TBE virus; however, the transmission mode of TBE virus in this tick population has not been investigated. This study was conducted in Białowieża Primeval Forest, east Poland. Forty fully engorged nymphs of D. reticulatus were sampled from root voles (Microtus oeconomus). Ticks were kept until molting. All ticks were screened for the presence of TBE virus by nested RT-PCR. Three adult ticks were positive for infection with TBE virus. The present study for the first time demonstrates the possibility of transstadial mode of TBEV transmission in D. reticulatus ticks.