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Valid measurement of substance use is necessary to evaluate preventive and treatment interventions. Self-report is fast and inexpensive, but its accuracy can be hampered by social desirability bias and imperfect recall. We examined the agreement between self-report of recent use and rapid diagnostic tests for three substances (alcohol, cannabis, and methamphetamine) among 904 young men living in Cape Town, South Africa. Rapid diagnostic tests detected the respective substances in 32, 52, and 22% of men. Among those who tested positive, 61% (95% CI [56%, 66%]), 70% ([67%, 74%]), and 48% ([42%, 54%]) admitted use. Men were moderately more willing to admit use of cannabis than alcohol (log OR 0.42) or admit use of alcohol than methamphetamine (log OR 0.53). Our findings show that self-report has reasonable criterion validity in this population, but criterion validity can vary substantially depending on the substance. [ABSTRACT FROM AUTHOR]