The Female Genital Tract Microbiome in Women with Endometriosis

In this study, we analyzed the microbiome composition of the reproductive tract - the vagina, endometrium, and pelvic peritoneum - in women with and without EMS using 16S rRNA gene sequencing.A total of 45 premenopausal women (20-45 years) with regular menstrual cycles were enrolled, including patients with surgically confirmed endometriosis (EMS) and controls undergoing surgery for dermoid cysts without evidence of EMS or adenomyosis. After quality filtering, eligible samples were included according to predefined exclusion criteria, including recent hormone therapy, probiotic or antibiotic use, prior abdominal surgery, pelvic inflammatory disease, pregnancy, malignancy, autoimmune disease, or BMI >30 kg/m2.Vaginal and endometrial specimens were collected under sterile conditions prior to surgery using OMNIgene vaginal collection devices, with strict precautions to avoid contamination. Samples were immediately stored at -80 C until DNA extraction.Microbial profiling was performed using 16S rRNA gene sequencing targeting the V3-V4 regions. Libraries were prepared according to the Illumina 16S Metagenomic Sequencing Library protocol and amplified using universal primers with Illumina adapter overhang sequences, followed by PCR under standard cycling conditions.