Objective: To study the relationship between the lactonase activities and status of paraoxonase 1 (PON1) and its association with the PON1 genetic polymorphisms in women with polycystic ovarian syndrome (PCOS). Design: A case-control study. Methods: A total of 455 PCOS patients and 441 control women were included in this study. The lactonase activities and concentrations of PON1 were assayed using 5-thiobutyl butyrolactone (TBBL) and 7-O-diethylphosphoryl-3-cyano-4-methyl-7-hydroxycoumarin (DEPCyMC) respectively. A normalized lactonase activity (NLA) was estimated based on the ratio of TBBLase:DEPCyMCase activity. The PON1 genotypes, serum malondialdehyde (MDA) levels and total antioxidant capacity were analyzed. Results: The lactonase activities and levels of PON1 were higher in PCOS patients than in the control women. However, the NLA did not significantly differ between groups. The K108C/T variation of the PON1 gene showed decreased lactonase activities and levels of PON1 in a genotype-dependent manner (CCOCTOTT); the 192Q/R variation of the PON1 gene showed increased PON1 lactonase activities and NLA; and the 55L/M variation of the PON1 gene showed decreased lactonase activities and levels of PON1 but an increased NLA. A multivariable regression analysis showed that the K108C/T, 192Q/R, and 55L/M variations of the PON1 gene, serum apolipoprotein A1, and MDA levels were significant predictors of PON1 lactonase activity, PON1 level, and NLA. Conclusions: The serum lactonase activities and concentrations of PON1 are increased in PCOS patients. The increased oxidative stress and the K108C/T, 192Q/R, and 55L/M genetic polymorphisms of PON1 may be associated with these changes.