Approaches for studying antimicrobial susceptibility and discovering new antimicrobial agents from the plants and other natural sources have been extensively utilized. Withania somnifera (L.) Dunal, commonly known as Ashwagandha or Indian ginseng or winter cherry, is a popular medicinal plant in Ayurvedic medicine. The principal active compounds include several withanolide-type compounds. Various plant parts, like roots and less often leaves and fruits of Ashwagandha, have been used as plant-derived medicines. The plant possesses various pharmacological activities including antimicrobial activity. Many bacterial and fungal species have been used as a test microorganism for the assessment of the antimicrobial activity of extracts and purified compounds of various plant parts of Ashwagandha. In this article, we tried to compile and to discuss the information about the antimicrobial activity of W. somnifera. This will provide the platform for the researchers to select plants, plant parts, solvent system, test microorganisms, method of evaluation and other related factors affecting the analysis.