The techniques of reverse transcription-polymerase chain reaction (RT-PCR) and restriction analysis were used to differentiate highly virulent Indian field isolates of infectious bursal disease virus (IBDV) from vaccine strains. Primers were designed to amplify the variable region of VP2 gene coding for major virus neutralizing epitopes. The 552 bp PCR products generated from four vaccine strains and five field isolates were digested with restriction enzymes DraI, HhaI, MvaI, StuI, StyI, and TaqI, which could differentiate field isolates from vaccine strains. Based on restriction enzyme profiles derived from published sequences, Indian field isolates seem to be closely related to highly virulent Japanese, European, and Chinese strains of the virus.