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Purpose: Self-reported behavior has been the cornerstone of measurement in HIV/STD prevention research. To evaluate the accuracy of self-reported sexual behavior, we evaluated selfreported abstinence following participation in an STD/HIV prevention trial and adolescents' acquisition of an incident STD at 6-month intervals over a two-year period post-intervention (i.e., 6-, 12-, 18-and 24-month follow-up assessments). Methods: Data assessing sexual behaviors were collected via self-administered ACASI interviews from 701 African American adolescent females (14-20 years, mean ϭ 17.67 years) prior to participating in a STD/HIV prevention trial. Additionally, adolescents provided self-collected vaginal swabs to assess the prevalence of three STDs. Participants were re-assessed (both selfreport interview and STD testing) at 6-, 12-, 18-and 24-months post-intervention. STD-positive adolescents were treated and received counseling at each time point. Contingency table analyses evaluated concordance between self-reported abstinence over each 6-month follow-up period and STD incidence separately and cumulatively across the 2-year follow-up period. Results: At each follow-up time point approximately 3 to 5% of participants reported abstaining from vaginal sex in the prior 6 months. However, among those reporting abstinence from vaginal sex, 25%, 27.3%, 22.2%, and 6.7% tested positive for an STD at the 6-, 12-, 18-, and 24-month follow-up assessment, respectively. Cumulatively, across the 2-year follow-up period, 77 sixmonth intervals of self-reported abstinence were reported, and 19.5% of individuals with a self-reported abstinence interval tested positive for a laboratory-confirmed STD. Further, among individuals who reported sexual activity within the same timeframe, approximately 20% tested positive for a laboratory-confirmed STD. Conclusions: Marked discordance was observed between adolescents' self-report of abstinence and their laboratory-confirmed STD incidence. These findings highlight the need to include biological markers to complement adolescents' selfreported sexual behavior and provide an objective and quantifiable marker for evaluating STD/HIV interventions.