In the present investigation, the methanolic and aqueous extract of roots of Withania somnifera were evaluated for antimicrobial activity against common human pathogens. It was observed that aqueous extract of Withania somnifera was more effective in inhibiting all the five test pathogens with zone of inhibition ranging between 33 mm and 50 mm as compared to methanolic root extract (15 to 38mm). The results indicate that the methanolic and aqueous root extracts of W. somnifera might be exploited as natural drug for the treatment of several infectious diseases caused by these organisms.