Hepatitis B remains a national health issue in Ghana. The prevalence of Hepatitis B varies widely among communities. Hepatitis B can prevail and spread through food vending activities. However, very scanty data is available on the subject. This study, therefore, sought to examine the prevalence of the Hepatitis B virus among food vendors in Kwadaso municipality. Two hundred and fourteen food vendors were screened for this exercise using the Hepatitis B Surface Antigen test. The test results were examined at a Diagnostic Laboratory. Primary data were generated through questionnaires and observations. There was a 9.3% prevalence of Hepatitis B Surface Antigen among food vendors in Kwadaso municipality. The ‘31–45’ age group had the highest prevalence of 40%. The Hepatitis B virus prevailed more in females (70%) than males (30%). One hundred and fifty-nine (74%) of the screened vendors were aware of their Hepatitis B status. Only one hundred and forty-eight (69%) of the study population screened on yearly basis. Majority (73%) of the vendors were roadside sellers. Most food vendors had high patronage of serving more than 200 customers daily. None of the food vendors had been vaccinated against the Hepatitis B Virus infection. Food, water, personal and environmental hygiene at food vendor premises must be closely monitored to ensure food security in the municipality. This is because there are poor sanitation practices in most of the food premises. The study therefore recommends more screening and vaccination exercises, health education, monitoring and food hygiene promoting programs as well as law enforcement initiatives on food vending activities to address the issue otherwise an outbreak of Hepatitis B virus infection through the food vending industry is likely to occur.