Background Since the beginning of the novel coronavirus disease in Wuhan city of China in 2019 and its spreading worldwide and taking the form of a pandemic, many healthcare workers (HCWs)were affected by coronavirus disease 2019 (COVID-19) infection. Though we have used many types of personal protective equipment(PPE) kitswhile taking care of COVID-19 patients, we have seen COVID-19 susceptibility in different working areas were different. The pattern of infection in different working areas depended on HCWsfollowing COVID-19 appropriate behavior. Therefore, we planned to estimate the susceptibility of front-line HCWs and second-line HCWs to getting COVID-19 infection. Aim To determine the risk of COVID-19 in front-line healthcare workers as compared to second-line healthcare workers. Method and materials We planned a retrospective cross-sectional analysis of COVID-19-positive healthcare workers from our institute within six months. Their nature of duty was analyzed and they were divided into two groups: 1) Front-line HCWs were defined as those who were working or who have worked in screening areas of the outpatient department (OPD)or COVID-19 isolation wards within the prior 14 days and provided direct care to patients with confirmed or suspected COVID-19. 2) Second-line HCWs were those who were working in the general OPD or non-COVID-19 areas of our hospital and did not have contact with COVID-19-positive patients. Results A total of 59 HCWs became COVID-19 positive during the study period, 23 as front-line and 36 as second-line HCWs. The mean (SD) duration of work as a front-line worker was 51 and as a second-line worker was 84.4 hours. Fever, cough, body ache, loss of taste, loose stools, palpitation, throat pain, vertigo, vomiting, lung disease, generalized weakness, breathing difficulty, loss of smell, headache, and running nose were present in 21 (35.6%), 15 (25.4%), 9 (15.3%), 10 (16.9%), 3 (5.1%), 5 (8.5%), 5 (8.5%), 1 (1.7%), 4 (6.8%), 2 (3.4%), 11 (18.6%), 4 (6.8%), 9 (15.3%), 6 (10.2%) and 3 (5.1%), respectively. To predict the risk of getting COVID-19 infection in HCWs, binary logistic regression with COVID-19 diagnosis as the output variable was modeled with hours of working in COVID-19 wards as front-line and second-line workers as independent variables. The results showed that there was a 1.18 times increased risk of acquiring the disease for every one-hour excess of working as a front-line worker, whereas, for second-line workers, it was slightly lower, with a 1.11 times increased risk for developing COVID-19 disease with every one hour increase in duty hours. Both these associations were statistically significant (p=0.001 for front-line and 0.006 for second-line HCWs). Conclusion COVID-19 has taught us the importance of COVID-19 appropriate behavior in preventing the spread of respiratory organisms. Our study has shown that both the front-line and second-line HCWs are at increased risk of getting the infection and proper use of a PPE kit or mask can decrease the spread of such respiratory pathogens.