The objective of the study was to assess the knowledge and perception of COVID-19 and relevant universal safety measures among the Nepalese population. A web-based cross-sectional study was conducted among Nepalese adults from March 29 to April 07, 2020. A 13- and 15- items structured questionnaire assessed the COVID-19 related knowledge and perception of the universal safety measure. Kruskal–Wallis test and Mann–Whitney U test evaluated the differences in knowledge between the groups. Data analysis was performed using IBM SPSS Statistics for Windows Version 21.0 (IBM Corp. Armonk, NY, USA). Of the 884 surveys accessed, a total of 871 consented (electronically) and completed the online survey (response rate 98.52%). The median knowledge score of the participants was 10.0 (± 3.0 IQR). Although participants’ overall knowledge score was high, only about half of the participants knew about the concept of quarantine and the ideal distance to be maintained between individuals to prevent the transmission. Though the majority of the participants had positive perception towards universal safety measure of COVID-19, about 18% perceived that coronavirus infected only older people, 11% opined that the infection was highly fatal with no chances of survival and 70% considered that limiting consumptions of poultry and meat would prevent the spread of COVID-19. A statistically significant difference in knowledge was noted by participants’ age, educational status, occupational type, and household monthly income. This study found optimal knowledge and perception of universal safety measures of COVID-19 among the Nepalese population, but misinformation and misunderstanding prevailed.