The clinical evaluation of female sexual dysfunctions should be supplemented by validated questionnaires; however, there is no specific instrument available in Greek language. The study was designed to linguistically validate the Greek version of Female Sexual Function Index (FSFI). Ninety-nine healthy women and eighteen women with a sexual dysfunction were recruited through a survey and were asked to voluntarily complete the FSFI questionnaire in Greek (FSFI-Gr) at baseline and after 2 weeks. We assessed validity, internal consistency reliability and test-retest reliability of the FSFI greek version. Subscales of the FSFI showed good internal consistency reliability (Cronbach's= 0.92, P<0.01). Test-retest reliability median 14 days apart was excellent with intraclass coefficient 0.91 (P<0.01). The FSFI-Gr discriminated between women with and without sexual dysfunction with an optimal cutoff score 26 (sensitivity 71.4%, specificity 92.2%). The Greek version of the FSFI is a reliable tool for the assessment of female sexual dysfunction. The results show that it is comparable to the outcome of studies in English-speaking countries.