Rabies is widespread in all provinces of Iran. This study presents an investigation by a Polymerase Chain Reaction (PCR) technique for the purpose of rabies diagnosis and also Restriction Fragment Length Polymorphism (RFLP) for determination of the type of the rabies virus. In addition, PCR proves to be a powerful tool for molecular epidemiology studies allowing analysis of the infecting strains without prior cell culture adaptation. In this study, the type of rabies viruses isolated from 50 brain samples of various hosts from different provinces of the country was determined by restriction analysis (RFLP) on pseudogene ? fragment as the most divergent genomic area. The results indicate that the viral strains belong to serotype/genotype 1.