<h3>Background</h3> The re-emergence of venereal syphilis cases is a public health concern, particularly in people living with HIV (PLWH). There is conflicting evidence whether or not Treponema pallidum (Tp) burdens are higher in early syphilis (ES) patients with HIV. The aim of this retrospective study was therefore to compare Tp burdens in blood and tissues of PLWH and people without HIV (PWH). <h3>Methods</h3> Patients with ES were consented and enrolled in Cali, Colombia between 2003 and 2020. Age, sex, HIV status and Tp polA results were extracted from available clinical and laboratory records. Data from PLWH and PWH were compared using Fisher's exact test for qualitative variables and Wilcoxon rank-sum test for age at time of diagnosis. The natural logarithm of polA copies/uL was regressed on HIV status and polA geometric mean ratios (GMR) and corresponding 95% confidence intervals (95%CI) were generated. <h3>Results</h3> Of 188 subjects with ES, 22 PLWH and 72 PWH had available polA results in blood or tissues. PLWH were more likely to be male (84% vs 50.6%, p=0.005). There were no differences in age (range 18–62) or ethnicity between PLWH and PWH. There were no differences in the proportion of patients with polA positivity between groups in blood (72% vs 55%, p=0.269), skin (64% vs 57%, p=0.763) or mucous membranes (67% vs 67%, p=1.000). There were also no differences in the geometric means of polA copies in the blood (GMR=1.8, 95%CI 0.5–6.0, p-value=0.339), skin (GMR=2.8, 95%CI 0.4–21, p-value=0.307) or mucous membrane samples (0.001, 95%CI 0–27, p=0.284). <h3>Conclusions</h3> HIV status was not associated with higher polA Tp burdens in blood or skin in subjects with early syphilis.