Herpes simplex virus (HSV) infection is a major opportunistic infection in immunosuppressed persons. The development of resistant strains of HSV to the available drugs for infection management, as is evident in the first drug of choice acyclovir, has further compounded this situation. There is therefore an urgent need to identify and develop new alternative agents for management of HSV infections, more so, for those due to resistant strains. We report here on methanolic extract preparation from the leaves of Hyssopus officinalis, a medicinal plant locally growing in Iran that has exhibited remarkable anti-HSV activity in vitro and in vivo for both wild type and resistant strains of HSV.