Tuberculosis (TB) is a pathogenic disease that affects a range of wildlife species, including African elephants (Loxodonta africana). The recent discovery of fatal disease caused by infection with Mycobacterium tuberculosis in a bull elephant in the Kruger National Park (KNP), which is a bovine TB endemic area, emphasizes the importance this disease could have on both wild and captive elephant populations globally. Elephants with culture-confirmed TB have previously been shown to produce strong antibody-responses before the mycobacteria can be isolated. Therefore, we used two serologic assays that detect TB antibodies to retrospectively screen a cohort of 222 free-ranging African elephants sampled between 2004 and 2018 in KNP. The estimated TB seroprevalence for this free-roaming elephant population was between 6% (95% confidence interval [CI], 2–12%) and 9% (95% CI, 6–15%) based on the two tests. Overall, males had a higher TB seroprevalence than females, and adults (≥25 yr) had a higher TB seroprevalence than younger elephants (≤24 yr) on both rapid tests. The relatively high TB seroprevalence that we found highlighted the value of conducting retrospective studies in free-ranging wildlife populations in order to better understand the potential risk of disease.