Purpose:  To access knowledge, attitude, and practices of Eye care practitioners (ECPs) towards COVID-19 guidelines and to determine their knowledge about use of Personal Protective Equipment (PPE). Methods:  A cross-sectional study was conducted in Tertiary eye hospitals of Rawalpindi and Islamabad, from October 2020 to March 2021. A total of 270 Eye care practitioners currently working in public and private hospitals were included in this study. Non-probability convenient sampling technique was used. Data was collected from individuals through an interview-based questionnaire after taking consent. Data was coded and analyzed by using SPSS version 26. The Chi-square test was used for further analysis. Results:  Mean age of participants was 29.85 ± 7.482 years, ranging from 18 to 65 years. Females (67%) were slightly outnumbered than males of the total sample. Moreover, 50% of the respondents had adequate knowledge about COVID-19 while only 39.3% had a positive attitude towards COVID-19. Hours of practice and years of experience had an association with knowledge. Good practices were observed by only 43.3% of the total sample. Practices had an association with the department of work of ECPs. Conclusion:  There is scarcity of knowledge among ECPs about COVID-19 and its preventive measures. Less than 50% ECPs with positive attitude and good practices towards COVID-19 indicate a dire need to train them for use of PPEs.