Objectives: The aim of this study was to assess and compare (a) the knowledge, attitude, and practice of standard precautions (SPs), (b) the knowledge of post-exposure management, and (c) the perceived barriers underlying the noncompliance with SPs among future healthcare professionals (HCPs), i.e., students of medical and nursing courses in Central India. Setting and participants: A cross-sectional study was conducted in 2017–2018 among students of a medical and a nursing college using a pretested and modified questionnaire. Data were collected during 23 face-to-face sessions. Responses were scored according to standard guidelines of the Centers for Disease Control and Prevention and WHO, where each correct response was given a score of 1. Results: Among 600 participants, 51% of medical students and 75% of nursing students could not select the correct definition of SPs from the given options. Sixty-five percent of medical students (275/423) and 82% of nursing students (145/177) were unaware of the term post-exposure prophylaxis. Overall, knowledge about personal protective equipment and hazard symbols was poor (<25%). Furthermore, although theoretical knowledge about hand hygiene was good (510/600; 85%), its implementation was poor (<30%). Sixty-four percent of participants believed that the use of hand rub replaced the need for handwashing, even for visibly soiled hands. Some of the participants believed that the use of PPE might offend patients (16%). High workload and poor knowledge were other significant barriers underlying the noncompliance with SPs. Conclusions: A suboptimal translation of participants’ knowledge into practice is evident and signifies the presence of the know–do gap. Poor knowledge and inappropriate presumptions about the use of SPs discourage the practice of SPs. This results in increased healthcare-associated infections, increased treatment costs, and a suppressed social economy. The inclusion of a dedicated curriculum with repeated hands-on and practice-based training on SPs is suggested to minimize this know–do gap among future healthcare workers.