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Objective: To determine the effect of genital hygiene behavior (GHB) and vaginal douching (VD) of pregnant women on preterm labor (PTL)., Methods: This study, which was conducted with 365 pregnant women, is a cross-sectional study. Data were collected using a questionnaire and the Genital Hygiene Behavior Inventory (GHBI) was applied., Results: The ratios of PTL diagnosis and VD practice, and mean GHBI scores of the pregnant women were 35.1%, 9.6%, and 64.42 ± 10.44, respectively. The prevalence of receiving a diagnosis of PTL was significantly higher in those who changed their underwear two to three times a week, took baths while sitting, cleansed their perineum with water using their hands, removed pubic hair at intervals of 2 months or longer, and performed VD. It was found that poor GHB and VD practices increased the rate of PTL and, in particular, VD was found to increase the risk of PTL around eight-fold., Conclusion: These results can help pregnant women to understand the relationship between genital hygiene and PTL and show the importance of being educated by health professionals about this issue., (© 2023 International Federation of Gynecology and Obstetrics.)