Background: Endometriosis (EMS) is a chronic gynaecological disease. Exosomal miRNAs appear to be associated with the progression of EMS, which suggest potential circulating biomarkers in EMS.Methods: Differential centrifugation, illumina small RNA sequencing, bioinformatics analysis and qRT-PCR are used to compare the pelvic fluid of patients with a clinical diagnosis of EMS and matched controls.Results: Six miRNAs (miR-135a-5p, miR-196a-5p, miR-449b-5p, miR-4454, miR-4286, and miR-5100) showed significant differences in the EMS group in initial screening (log2FC > 2). Receiver-operator curve (ROC) analysis further indicated miR-4454 (area under the curve (AUC) = 0.956, P < 0.05) and miR-5100(area under the curve (AUC) = 0.900, P < 0.05) were highly correlated with the pathogenesis of EMS. Validation on the samples from 10 patients and 10 healthy people.Conclusion: Our miRNA expression data provide altered expression and potential prospects for biomarkers in EMS.