Over the past decade, Hospital-acquired infections (HAIs) have become a significant concern within Indian Tertiary Healthcare Centers. The effectiveness of infection control practices heavily relies on the diligence of healthcare workers, directly impacting the success of these measures. It's crucial to closely examine any disparities between ideal and actual practices to ensure optimal infection control outcomes. To find any gap between ideal and actual infection control practices and analyze associated factors which is actually responsible for the that gap. A cross-sectional study was carried out at a tertiary healthcare center in north Gujarat over six months, spanning from February 2023 to July 2023. The study included a total of 293 participants, comprising 61 doctors, 154 nurses, 33 paramedical staff, and 45 housekeeping staff. Data were collected using a pretested, structured, self-administered questionnaire covering different aspects of infection control practices. Knowledge, attitude, and practice (KAP)Scores of 70% or less were considered below average, scores between 71 and 80% average, 81 to 90% good and above 90% were very good. KAP score is 98.8% in knowledge, 88.70% in attitude and 89% in self-reported practices. Cumulative KAP score was 92.16% which is under very good category. Around 88.52% (54 out of 61) of doctors, 69.48% (107 out of 154) of nurses, 78.78% (26 out of 33) of paramedical staff, 64.44% (29 out of 45) of housekeeping staff scored above 90% in KAP score which is under very good category.Regular training, post training assessments and strict enforcement of infection control measures in hospitals can help to fill gaps in the knowledge, attitudes, and self-reported practices of healthcare workers regarding infection control.