Melioidosis is a disease of public health importance associated with high case-fatality rates in animals and humans caused by Burkholderia pseudomallei (B. pseudomallei), a gramnegative bacterium that lives in tropical soil environments. This study determined the seropositivity for melioidosis among patients admitted to healthcare centres in Malaysia, from 2015 to 2019 and identified factors related to it. A total of 26,665 serum samples of suspected melioidosis patients from Malaysia hospitals were tested for IgM against B. pseudomallei. About 16.4% of total samples were seropositive and majority of them were 55 years old and above. However, younger people aged less than 15 years old were the most susceptible to the infection (AOR 3.04, p <0.001, 95% CI: 2.73, 3.39). Melioidosis infection was the highest in Sarawak (15.1%) while Perlis was the least exposed to the infection (0.9%). Further analyses showed that patients with chronic lung disease (adjusted OR: 4.03, p < 0.001, 95% CI: 2.77, 5.86) were more susceptible to melioidosis infection. In conclusion, although serology testing is not a gold standard test in diagnosing melioidosis, it has been used as a tool in treatment monitoring and disease surveillance among patients and at-risk community in the endemic hot-spots regions.