This study investigated the expression profiles of two specific microRNAs (miRNAs), miR-1271-5p and miR-10a-3p, in patients with bacterial vaginosis (BV) and in healthy controls. One hundred participants (50 patients with BV and 50 controls) were recruited. Bacterial species were identified using multiplex PCR of the 16S rRNA gene from vaginal swabs, and miRNA expression was quantified from blood samples using RT-qPCR. Gardnerella vaginalis was the most prevalent species (66.0%) in patients with BV. The expression levels of miR-1271-5p and miR-10a-3p were significantly higher in patients with BV than in healthy controls (p < 0.001). Receiver operating characteristic curves revealed promising diagnostic values: an miR-1271-5p cutoff of <5.42-fold achieved 94.0% sensitivity and specificity (area under the curve (AUC), 0.951), whereas an miR-10a-3p cutoff of <3.57-fold showed 96.0% sensitivity and 92.0% specificity (AUC, 0.970). Interestingly, miR-1271-5p and miR-10a-3p levels were the highest in patients with Megasphaera colonization. Thus, miR-1271-5p and miR-10a-3p are potential biomarkers for the diagnosis of BV and offer high diagnostic value. These miRNAs may play a crucial role in understanding BV pathogenesis and serve as targets for future diagnostic strategies.