This prospective cohort study was conducted at a tertiary medical center (IRB#0390-20-SOR). The study population included pregnant women with a history of spontaneous singleton preterm birth (PTB). Participants provided saliva samples and completed validated stress questionnaires twice during pregnancy. The incidence of rPTB was compared between women with high versus low cortisol levels and perceived stress, defined as greater than the 75th percentile for the cortisol levels and the questionnaire score. Multivariable logistic regression models were used to assess the association between elevated stress markers and the risk of rPTB.