We aimed to evaluate the knowledge about chlamydial infection of Greek midwives and midwifery students. An appropriately designed, self-administered, anonymous questionnaire was distributed to 107 midwives and 29 graduating midwifery students. Perceived awareness of chlamydial infection was similar in midwives and students (p=0.083). However, midwives were more aware than students that Chlamydia are acquired through sexual contact (84.1% vs 58.7%; p=0.004) and that chlamydial infection is frequently asymptomatic (72.9% vs 37.9%; p=0.001). Knowledge of the consequences was poor in both groups, but it was relatively better in midwives; only 7.5% of the midwives did not know any of the potential sequelae of chlamydial infection compared with 24.1% of the students (p=0.006). In contrast, the majority of both midwives and students were aware that when chlamydial infection is diagnosed in a woman, both she and her partner should receive treatment (90.7% vs 93.2%; p = 0.901). In conclusion, an important proportion of midwifery students ignore important aspects of chlamydial infection. Therefore, it is important to improve the educational curriculum in midwifery schools in order to maximize the contribution of midwives to decrease the prevalence of chlamydial infection and of its complications.