Objectives <p>Integrating standard treatment guidelines (STGs) into clinical practices is crucial for improved healthcare quality and patient safety. This study explores clinicians' awareness, attitudes, and perception of barriers and facilitators for adoption of STGs in clinical practice.</p> Methods <p>A cross-sectional prospective study was conducted in two parts. The first part comprised of twelve online webinars that were conducted between 2021 and 2022 aimed at promoting STG awareness and their adoption. These webinars utilized case-based quizzes to highlight variations in disease management and encouraged discussions on their clinical impact. The second part comprised an online survey to assess the participants’ attitudes and practices, reasons for not using STGs, and barriers and facilitators for its effective adoption of the guidelines in clinical practice. Descriptive statistics (frequency and percentage) were used to describe the study subjects. Correlation analysis was also done to explore the relationship between two variables and was used to find out the significance of intervention (<i>P</i> &lt; 0.05) about the knowledge and practice of the study subjects.</p> Results <p>Out of a total of 1300 cumulative attendees in the webinar, 355 physicians participated in the quizzes and 260 physicians undertook online survey on awareness, attitude, perceived barriers and facilitators for guideline implementation respectively. Quizzes during the session revealed significant clinical variability (particularly in mother and child health, role of prophylaxis and prevention of infection diseases, management of ADRs screening &amp; threshold criteria for assessment of diabetes, hypertension etc., indicating that past practices and personal experience superseded standardized approaches. The survey indicates that personal experience (26.9%) is the predominant factor in clinical decision-making, followed by clinical guidelines (18.1%), peer-reviewed research articles (15.8%), and discussions with peers/colleagues (12.3%). Notably, 60.8% of respondents cited difficulty in making definitive diagnoses, and 25% reported a lack of awareness about existing guidelines as major reasons for not utilizing clinical guidelines. The other obstacles included limited access to guidelines, misconceptions about legal implications, and reimbursement restrictions. Potential facilitators for STG adoption were clear guidelines, regular audits, comprehensive training, and digital integration, as well as guideline adherence and performance incentives.</p> Conclusions <p>The findings underscore the necessity of guidelines for standardizing care and ensuring patient safety. To enhance STG adoption, there is a need for improved access to clear guidelines, sensitization regarding their importance guideline adherence, and addressing legal misconceptions. Sustainable change requires educating younger physicians and integrating STGs into medical curricula. India should adopt enforcement mechanisms like reimbursement schemes and make guideline adherence a requirement for accreditation to foster a culture of guideline-based practice.</p>