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Currently, there are no validated instruments in Italian specifically designed to assess mental illness stigma or prejudice. Moreover, implicit measures, while insightful, are often resource-intensive and impractical for large-scale population studies of Italian speakers. The present study investigated the validity of the Italian version of the Prejudice towards People with Mental Illness scale (PPMI-IT) in measuring biases toward individuals with mental health issues. The original instrument by Kenny et al. was translated from English into Italian and vice versa. A sample of 455 Italian-speaking participants (65% female; M_age = 33.39; SD = 13.21) was utilized to conduct a confirmatory factor analysis, confirming a four-factor structure (fear/avoidance, malevolence, authoritarianism, unpredictability). Factor loadings indicated that each dimension was well represented, supporting the construct validity of the scale. Model fit indices, including chi-square (χ² = 782.54, df = 296.00, χ²/df = 2.64), RMSEA (0.06, 90% CI: 0.060-0.07), CFI (0.93), TLI (0.91), and SRMR (0.06), suggest an excellent model fit. Furthermore, the analysis of correlations and the heterotrait/monotrait (HTMT) ratio provides evidence supporting the discriminant validity of the PPMI scale compared with social desirability. These findings confirm that the PPMI scale is a valid and reliable tool for assessing biases toward individuals with mental health issues, making it suitable for academic research, clinical interventions, and public policy contexts.