Abstract: OBJECTIVE To explore the relationship between genital tract mycoplasma infection and premature rupture of membrane (PROM) and observe its influence on mitochondrial apoptosis pathway. METHODS A total of 121 puerpera with genital tract mycoplasma infection and 118 puerpera without genital tract infection who gave birth in Yuncheng Nursing Vocational College from Dec 2019 to Dec 2021 were recruited as the research subjects and were respectively assigned as the infection group and the healthy group. The incidence of PROM and pregnancy outcomes were compared between the two groups. The puerpera of the infection group were divided into the PROM group with 45 cases and the non-PROM group with 76 cases according to the status of PROM. The levels of cysteine proteinase-3 (Caspase-3), Bcl-2 associated X protein (Bax) and B-cell lymphoma/leukemia-2 (Bcl-2) in fetal membrane samples were detected by means of enzyme-linked immunosorbent assay. The levels of fetal membrane tissue Caspase-3, Bax and Bcl-2 were compared between the two groups, and the values of fetal membrane tissue Caspase-3, Bax and Bcl-2 in prediction of genital tract mycoplasma infection and PROM were analyzed by receiver operating characteristic (ROC) curve. RESULTS The proportion of the puerpera with PROM, preterm delivery, neonatal pneumonia and pathologic jaundice were higher in the infection group than in the healthy group (P<0.05). The levels of fetal membrane tissue Caspase-3 and Bax of the infection group were higher than those of the healthy group(P<0.05). The levels of fetal membrane tissue Caspase-3 and Bax of the PROM group were higher than those of the non-PROM group(P<0.05), the Bcl-2 level of the PROM group was lower than that of the non-PROM group(P<0.05). The area under curves (AUC) of Caspase-3, Bax, Bcl-2 and the joint detection were respectively 0.706, 0.741, 0.768 and 0.914, and the AUC of the joint detection was larger than that of the single detection(P<0.05). CONLCLUSIONS The genital tract mycoplasma infection may increase the risk of PROM and adverse neonatal outcomes, which may be associated with the activation of mitochondria-dependent apoptosis pathway in fetal membrane tissues.