Background: Dengue, a major global public health concern, is growing more prevalent. Extensive measures have been undertaken to prevent and control dengue transmission in Malaysia including understanding and modification of human behaviour. This study was designed to assist in planning an effective health intervention for dengue by measuring residents’ knowledge, attitudes, and practice (KAP) in hotspot areas of Sabah, East Malaysia. Methods: A community-based and cross-sectional study was conducted at dengue hotspot area in Sabah involving 463 residents (aged 18 years and above). Information on the socio-demographic characteristics of the residents and their KAP towards dengue was collected using a structured pre-validated questionnaire developed by the researchers. In addition, residents were invited to participate in a dengue seroprevalence study using dengue Immunoglobulin M (IgM) and Immunoglobulin G (IgG) ELISA kit. Results: Most of the residents correctly identified common signs and symptoms of dengue such as fever (84.4%), chills (70.7%), and nausea or vomiting (56.0%). Nearly half of the residents (51.3%) had good knowledge, 66.0% had a positive attitude, and 63.7% had a positive practice for dengue prevention and control, respectively. The dengue seroprevalence study included 200 of the 463 residents; IgG and IgM seropositivity was 74.1% (n = 63) and 7.1% (n = 6) respectively. Based on logistic regression analysis, local and migrants differs considerably in terms of their knowledge and attitude toward dengue prevention and control, while gender and level of education were predictive of practice level (p<0.05) among all sociodemographic variables examined. Knowledge was correlated with attitude (p<0.0001) and practice (p< 0.0014). Discussion: To combat dengue epidemics in the hotspot area of Sabah, a proactive and behavioural changes among residents, are required. To effectively prevent and manage dengue transmission, a collaborative effort of government sectors, private sectors, housing residential committees, and community residents is essential.