Youth living with HIV (YLWH; aged 16-24) are at elevated risk of alcohol and drug use. Studies in older populations have identified patterns or profiles of multiple substance use differentially associated with mental health and anti-retroviral therapy (ART) adherence. No studies of YLWH have yet examined such patterns. A sample of 179 YLWH, reporting ART non-adherence and alcohol use, were recruited at five Adolescent Trials Network clinics in urban areas of the US between November 2014 and August 2017. Participants completed the Alcohol Smoking and Substance Involvement Screening Test (ASSIST) to assess substance use involvement scores, and the Brief Symptom Inventory. Latent Profile Analysis identified three substance use patterns: minimal illicit drug use (15.1%), cannabis only (56.4%), and global polysubstance use (28.5%). Global polysubstance users experienced more mental health problems compared to the minimal illicit drug use group. The co-occurrence of drug use with alcohol was common among these YLWH-all of whom reported ART adherence problems-indicating the importance of interventions capable of addressing multiple substance use rather than alcohol alone.