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One of the fast-growing major non-communicable diseases (NCD) that poses a danger to global public health is Diabetes mellitus (DM). Trends in the incidence of DM indicate a disproportionate increase in developing countries due to current rapid demographic transitions from traditional to more westernized and urbanized lifestyles. Knowledge of DM is vital for curbing or control. The objectives of this study were to evaluate the level of knowledge and awareness of DM among the Ho municipality general population, identify areas of deficiency for targeted health education efforts, and identify respondent characteristics that may be associated with knowledge of diabetes. A survey involving 132 respondents (age over 18 years) was conducted in the Ho municipality of the Volta region of Ghana. A 42-item pre-tested questionnaire was administered to participants to evaluate general and specific knowledge and awareness of DM. The Pairwise Multiple Comparison and Fisher’s Exact tests were used to test the hypotheses and associations between the respondents’ knowledge level and groups respectively. Of the 132 respondents, 22% were in the age range of 40-46 years; 72.7% were female. Mean over all diabetes knowledge composite score was poor: 32.99% (CI; 27.5, 38.5). Respondents performed best in the symptoms section: mean score was 36.247% (CI; 29.0, 43.4); and worst in the section on complications: mean score was 30.909% (CI; 23.6, 38.2). In multiple linear regression analyses, education level, older age, own self having diabetes, and having a family member/relative/friend with diabetes were significantly associated with knowledge of diabetes. Knowledge of diabetes among the inhabitants of Ho municipality respondents was interpreted as being inadequate 32.99% (CI; 27.5, 38.5). Some deficient portions and factors associated with knowledge of diabetes were identified. Relevant information for targeted health education programs in Ghana and beyond may be considered as one of such benefits of these findings.