Background: Treponemal infection is contagious and one of the oldest blood-borne infections, with great public health consequences. This study aimed to comparatively describe the five-year (2013-2017) regional epidemiology of Treponemal infection using pregnant women in the Sentinel Survey and apparently healthy blood donors as proxy for the general population at the four sentinel sites in the Volta Region of Ghana. Method: We analysed retrospectively data from 17,744 prospective blood donors aged between 18 to 58 years and 7,805 pregnant women in a Sentinel Survey who fell within the 15 and 49 years age bracket at Hohoe, Ho, Tongu and Krachi-West sentinel sites in the Volta Region. Data extracted included age, gender, date of blood donation and Treponena pallidum chromatographic immunoassay results from the blood banks of the four study sites.Published reports of Sentinel Surveys conducted at the four sentinel sites from the years 2013-2017 were retrieved. Results: The cumulative five-year prevalence of Treponemal infections among the pregnant women in the Sentinel Survey and prospective blood donors was 0.38% and 2.38% respectively. Site-specific prevalence for population-base/Sentinel survey was 4.6%/0.4%, 2.0%/0.2%, 1.3%/0.8 and 1.2%/0.2 for Hohoe, Ho, Krachi-West and Tongu respectively. Treponemal infection rates among the younger age groups (15-24years) were 0.31% in the sentinel survey and 2.22% in the general population. Significant gender disparity in Treponemal infection exist with male preponderance. Conclusion: The regional prevalence of Treponemal infection in the Sentinel Survey is lower compared to the general population. Therefore, the use of pregnant women as proxy for population estimate could lead to underestimation of the burden in the study jurisdiction.