Introduction Although only a few cases of monkeypox have been reported in India so far, it is vital for healthcare practitioners to have sufficient knowledge about its epidemiology, clinical presentation, diagnosis, and management. Therefore, this study aimed to understand the perception of Indian healthcare practitioners regarding the 2022 outbreak of monkeypox disease. Methods A cross-sectional survey was conducted among 500 medical and dental practitioners from various regions of Kerala, India. The Chi-squared test for proportion was used to determine the significant difference in the knowledge levels of the participants. Binary logistic regression (multivariate) was used to understand the knowledge of healthcare professionals regarding the 2022 outbreak of monkeypox disease. Results A total of 424 healthcare professionals completed this survey. Overall, the level of knowledge was good in 64.9% and poor in 35.1% of the participants (p<0.01). Binary logistic regression analysis did not show any statistical significance (p > 0.05) in terms of demographic characteristics. However, in this sample, participants with 6-10 years of work experience were more likely to have improved knowledge scores (odd's ratio OR 1.764). Similarly, participants between the age of 30-40 years were also likely to have improved knowledge scores (OR: 1.065). Conclusions Indian healthcare professionals had an overall good level of knowledge regarding the 2022 outbreak of monkeypox. However, a low level of knowledge was found related to the clinical presentation and immunization of monkeypox. This may be due to the low prevalence of monkeypoxin this region.