Background As a clinical and non-invasive tool, the AGE Reader measures skin autofluorescence (SAF) to estimate the accumulation of advanced glycation end products (AGEs) in the skin. Accumulation of AGEs has been implicated in several inflammation-associated diseases, including diabetes and cardio-metabolic diseases. This study aimed to assess SAF in subjects with and without cardiovascular risk (CVR) factors and examine the association between SAF and various bio-clinical parameters. Methods In a cross-sectional study, we included 250 participants between 19 and 86 years of age divided into two groups: a healthy group (n = 88) and subjects with CVR factors (n = 162 in total, diabetes n = 48, hypertension n = 62, and both n = 52). We assessed skin AGE measures and biological and clinical data.Results SAF was significantly higher in subjects with CVR factors than in healthy participants (2.42 ± 0.38 vs 1.90 ± 0.29 respectively; p < 0.001). SAF was associated with age, gender, BMI, duration of diabetes, HbA1c, triglyceride, and obesity. Multivariate analysis showed that age and duration of diabetes were the independent determinants of SAF. The ROC analysis indicated that a SAF > 2.25 AU was the optimal cut-off point to predict the presence of diabetes and/or hypertension and dyslipidemia (p < 0.001). Conclusion This Tunisian population-based study shows an increased SAF level in subjects with diabetes and/or hypertension and dyslipidemia compared to healthy subjects. The AGE Reader device is a rapid and non-invasive tool in clinical practice to evaluate and screen CVR factors in Tunisia with a North African phototype.