Experimental pain testing requires specific equipment and may be uncomfortable for patients. The Pain Sensitivity Questionnaire (PSQ) was developed to assess pain sensitivity, based on the pain intensity ratings (range: 0–10) of painful situations that occur in daily life. The main objective of this study was to carry out a cross-cultural adaptation and validation of the Spanish version of the PSQ (PSQ-S). A total of 354 subjects (296 healthy and 58 chronic pain patients) filled in the PSQ-S. A subgroup of 116 subjects performed experimental pain testing, including two modalities (cold and pressure), with different measures: pain intensity rating, pressure pain threshold, and tolerance. The validation results showed two factors: PSQ-S-moderate and PSQ-S-minor and, for the total scale and the two factors, an excellent internal consistency (Cronbach's alpha coefficient > 0.9) and a substantial reliability (Intraclass Correlation Coefficient > 0.8). We obtained strong correlations with all the experimental pain rating parameters, catastrophizing, and depression variables, as well as moderate correlations with anxiety, central sensibilization, and impact on the quality of life. Chronic pain patients received elevated PSQ-S scores compared to healthy controls, and three cut-off values (PSQ-S-total = 7.00, PSQ-S-moderate = 7.57, and PSQ-S-minor = 6.29) based on ROC curve analyses were shown to be able to discriminate between healthy adults and adults with chronic pain. Therefore, PSQ-S may be a simple alternative to experimental pain procedures for clinical and experimental pain research. [ABSTRACT FROM AUTHOR]