Background: With the increasing global movement of labor migrants, the risk of cross-border transmission of infectious diseases remains a public health concern. Pre-departure medical screening plays a vital role in identifying asymptomatic carriers and preventing disease spread. This study aimed to evaluate the seroprevalence of key infectious markers among pre-departure migrant workers and highlight implications for public health screening policies. Methods: This cross-sectional study was conducted at the Department of Biochemistry, Muscat Medical Center, Dhaka, Bangladesh, from January to December 2024. A total of 450 migrant workers undergoing routine pre-employment medical screening were enrolled. Blood samples were analyzed for HIV I/II, Hepatitis B surface antigen (HBsAg), Hepatitis C virus (HCV) and syphilis (VDRL and TPHA). Data were analyzed using SPSS version 25. Results: Among the 450 participants, the majority were male (92%) and aged between 20–39 years (82%). Most had completed at least primary (30%) or secondary education (42%). Regarding medical fitness, 93.8% were declared fit, 5.1% held up and 1.1% unfit. Only HBsAg showed seropositivity: 5 individuals (1.1%) tested positive, of whom 2 were unfit and 3 held up for confirmation. All participants tested negative for HIV, HCV, TPHA and VDRL. A detailed review showed 442 (98.2%) were HBsAg negative, 2 (0.4%) positive and 6 (1.3%) required retesting. Conclusion: The overall seroprevalence of infectious markers was low, with Hepatitis B being the only positive finding. These results support the importance of maintaining comprehensive pre-departure infectious disease screening to ensure public health safety and early intervention.