Background
There is growing scientific interest in characterizing the cervicovaginal microbiota (CVM) in the context of cervical cancer, particularly the role of specific bacterial species that may influence the persistence of human papillomavirus (HPV) and progression to high-grade lesions or invasive disease. It has been proposed that microbial profiles vary according to HPV genotype and infection status. In this study, we aimed to identify the most frequent non-HPV coinfections in HPV-positive and HPV-negative women and explore their potential role in shaping the cervicovaginal microenvironment
Objectives
To characterize the prevalence and distribution of non-HPV sexually transmitted pathogens in HPV-positive and HPV-negative women from underserved regions of Colombia using next-generation sequencing, and to assess their association with HPV genotypes, single and multiple infection patterns, and their potential role in modulating the cervicovaginal microenvironment and disease progression.
Methods
We analyzed 433 liquid-based cytology samples from women aged 30 to 65 years, from underserved and remote regions of Colombia (coastal, Andean-mountain, and Amazon areas), with diverse sociodemographic risk factors. Cytological findings were classified according to the Bethesda system. All samples were analyzed by next-generation sequencing (NGS) using the Chapter Diagnostics HPV and Non-HPV Sexually Transmitted Infection (STIs) kit on the Illumina MySeq platform. This allowed for the identification of 19 high-risk HPV genotypes, 10 low-risk genotypes, and 19 non-HPV pathogens associated with sexually transmitted infections (STIs).
Results
We identified 47 HPV infections (34 single and 13 multiple) and 48 samples with multiple HPV infections plus coinfections with non-HPV pathogens. Additionally, 125 infections exclusively by non-HPV pathogens were detected (97 single and 28 multiple). A total of 101 HPV genotypes were identified: 62 high-risk and 39 low-risk. Overall, 248 pathogens were detected, with Ureaplasma urealyticum, Ureaplasma parvum, and Trichomonas vaginalis being the most prevalent. HPV-positive samples predominantly showed multiple coinfections with U. parvum and Mycoplasma hominis, while HPV-negative samples mostly presented single infections, with Trichomonas vaginalis being more frequent.
Conclusion
Differences in microbial coinfections were identified between HPV-positive and HPV-negative women. Characterizing the cervicovaginal microbiota could enhance molecular surveillance and support early detection and prevention strategies for cervical cancer.