Two Herpes simplex viral serotypes, HSV type 1 and HSV type 2 may cause genital herpes. The aim of this study was to perform a genetical analysis and characterization of virus isolated from four patients with double genital herpetic infections. In eleven viral isolates, the cytopathic effect in Vero cells was studied, the antigenic type was determined using monoclonal antibodies and genomic analysis was performed with Eco RI, Hind III and Bgl II enzymes. Five viral isolates generated a diffuse and 6 a localized cytopathic effect. Monoclonal antibodies identified four HSV-1 and seven HSV-2. Genomic analysis had concordant results. Four HSV-1 were obtained, with different genomic patterns within them, three were different to the standard North American strain. The seven HSV-2 obtained had three different types of electrophoretic profiles, that were different to the standard North American strain. It is concluded that the genomic and antigenic analysis allowed the detection of herpetic genital infections caused by herpes virus type 1 and 2 in the same individual and the identification of herpes virus strains with distinct regional characteristics.