Human monkeypox (now termed as the “Mpox”) was considered a geographically isolated disease until recently, but the current spread of the disease over 110 locations around the world has created an environment of fear. This study was thus conducted to evaluate the knowledge and apprehensions about the ongoing outbreak of Monkeypox among the medical students and nursing staff who can be a vital source of dissemination of knowledge to the general population. A semi-structured, self-administered questionnaire, was used in this cross-sectional offline study. The study population comprised Medical undergraduate students and nursing staff of a medical college situated in northern India. A total of 340 participants took part in the study comprising 302(88.8%) medical undergraduates and 38(11.1%) nursing staff. Overall, the knowledge of medical students and nursing staff was unsatisfactory. When the questionnaire data were analyzed only 17.05% of the participants had good knowledge, 20.58% had moderate knowledge and 65.78% had poor knowledge. Overall knowledge about the human monkeypox virus and disease was underwhelming. Medical students and nursing staff can be a vital source of dissemination of knowledge to the general population. In this era of emerging threats; to fill the knowledge gaps of the health care professionals’ strategies like continuing medical education, webinars, seminars, and workshops primarily focussing on better clinical, prevention, and control practices should be conducted frequently.