This study aimed to evaluate the psychometric properties of TECO-D for Biomedical Laboratory Techniques students at Sapienza University of Rome. The study was conducted by the research group “Riabilitazione, Evidenza e Sviluppo” (R.E.S.) based at the same university. The sample included 494 students enrolled in the Bachelor’s degree program in Biomedical Laboratory Techniques, divided into four groups based on their academic year. TECO-D test consisted of 70 multiple-choice questions (MCQs) selected from a database of 250 questions and was administered through a digital platform. Psychometric analyses were performed using SPSS-23, evaluating internal consistency (Cronbach’s alpha), cross-cultural validity (two-way ANOVA), and discriminant validity (Receiver Operating Characteristic—ROC curve analysis). TECO-D demonstrated excellent internal consistency, with a Cronbach’s alpha of 0.889. Cross-cultural validity analysis revealed a statistically significant improvement in mean scores across different academic years, confirming the test’s ability to assess competence progression. Discriminant validity analysis showed an Area Under the Curve (AUC) of 0.85 (95% CI 0.81–0.88) for first-year students, 0.84 (95% CI 0.80–0.88) for second-year students, and 0.83 (95% CI 0.77–0.90) for third-year students, indicating a moderate ability to distinguish competency levels. TECO-D is a reliable and valid instrument for assessing competencies in Biomedical Laboratory Techniques students. It effectively tracks learning progression and competency development, making it a useful tool for improving educational quality and monitoring university training outcomes. This study supports the integration of TECO-D into academic programs as a standardized assessment method for evaluating students’ knowledge acquisition and professional readiness.