<p>In order to improve spontaneous preterm birth (sPTB) risk stratification in a predominantly white cohort of non-labouring pregnant women, we analysed their vaginal microbiota, metabolite, cytokine and foetal fibronectin (FFN) concentrations at two gestational time points (GTPs): GTP1 (20⁺⁰–22⁺⁶ weeks, preterm = 17; term = 32); and GTP2 (26⁺⁰–28⁺⁶ weeks, preterm = 14; term = 31). At GTP1, the preterm-delivered women showed abundant <i>G. vaginalis</i> (AUC = 0.77) over <i>L. crispatus</i> and <i>L. iners</i>, and upregulation of 10 metabolites. At GTP2, the same women had more lactobacilli- and mixed anaerobes-dominated microbiota, upregulation of five metabolites, and decreased TNFR1, distinguishing them from their term counterparts (AUC = 0.88). From GTP1 to GTP2, sPTB was associated with increased microbiota α-diversity, and upregulation of pantothenate and urate. CXCL10 declined in the term-delivered women by ~3-fold, but increased in the preterm-delivered women (AUC = 0.68), enhanced by FFN (AUC = 0.74). Characterising the complex dynamic interactions between cervicovaginal microbial metabolites and host immune responses could enhance sPTB risk stratification.</p>