The concurrent, construct, discriminative and predictive validity of the World Health Organization's Alcohol Substance Involvement Screening Test (ASSIST) were examined in an Australian sample. One hundred and fifty participants, recruited from drug treatment (n = 50) and primary health care (PHC) settings (n = 100), were administered a battery of instruments at baseline and a modified battery at 3 months. Measures included the ASSIST; the Addiction Severity Index-Lite (ASI-Lite); the Severity of Dependence Scale (SDS); the MINI International Neuropsychiatric Interview (MINI-Plus); the Rating of Injection Site Condition (RISC); the Drug Abuse Screening Test (DAST); the Alcohol Use Disorders Identification Test (AUDIT); the Revised Fagerstrom Tolerance Questionnaire (RTQ); and the Maudsely Addiction Profile (MAP). Concurrent validity was demonstrated by significant correlations between ASSIST scores and scores from the ASI-lite, SDS, AUDIT and DAST; and significantly greater ASSIST scores for those with diagnoses of abuse or dependence. Construct validity was established by significant correlations between ASSIST scores and measures of risk factors for the development of drug and alcohol problems. Participants diagnosed with attention deficit/hyperactivity disorder or antisocial personality disorder had significantly higher ASSIST scores than those not diagnosed as such. Discriminative validity was established by the capacity of the ASSIST to discriminate between substance use, abuse and dependence. ROC analysis was able to establish cut-off scores for an Australian sample, with suitable specificities and sensitivities for most substances. Predictive validity was demonstrated by similarity in ASSIST scores obtained at baseline and at follow-up. The findings demonstrated that the ASSIST is a valid screening test for psychoactive substance use in individuals who use a number of substances and have varying degrees of substance use.