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Although the etiology of cervical cancer has been partially established, its pathogenesis remains a key research focus. Specific studies on the genital tract microbiota of cervical cancer patients in Southern China are still scarce. To address this research gap, the present study collected genital tract microbial samples from 16 cervical cancer patients and 16 healthy women. Analysis of cervical cancer-specific microbiota was conducted via 16S rRNA gene sequencing. The study included 32 women (16 cervical cancer patients and 16 healthy controls) aged 45-65 at enrollment. Total genome DNA from samples was extracted using the hexadecyltrimethylammonium bromide (CTAB) CTAB method. The vaginal and cervical microbiota composition was determined by sequencing barcoded 16S rDNA gene fragments (V3-V4), and a comparative bioinformatics analysis of the microbiome was performed. The study revealed a homogeneous microbial composition dominated by Lactobacillus in healthy women, whereas cervical cancer patients exhibited increased diversity with reduced Lactobacillus and enriched Prevotella. No significant differences were observed between sampling sites within each group. Functional predictions linked the cancer-associated microbiota to metabolic pathways and the AEROBACTIN-SYN-PWY pathway. In conclusion, our findings suggest that these specific key microbial taxa and their related metabolic pathways contribute to the pathogenesis of cervical cancer and may serve as promising targets for clinical treatment and intervention.