While structured clinical interviews are considered the gold standard for diagnosing mental disorders, respective instruments are still lacking in the field of sexual dysfunctions. The study evaluates the psychometric properties of the new Diagnostic Interview for Sexual Dysfunctions in Women (DISEX-F), which is based on the eleventh edition of the International Statistical Classification of Diseases (ICD-11) and the fifth edition of the Diagnostic and Statistical Manual of Mental Disorders (DSM-5), in a sample of 100 women with self-reported sexual problems. Participants were interviewed twice by trained diagnosticians with the DISEX-F. A third diagnostician evaluated the audio records of the initial interview. Participants also completed self-report measures of sexual functioning/distress and interview acceptance. The DISEX-F demonstrates excellent inter-rater reliability, good test-retest reliability, and strong convergent and discriminant evidence of validity. Furthermore, it achieves high acceptance among participants. Discordant diagnostic outcomes were especially linked to false differential diagnostic decisions and information variance in participants reporting. The results strongly support using the DISEX-F for women presenting with self-reported sexual problems in practice and research.