A Correlation Study Between Cervical Microbiota And HR-HPV Infection Based On 16S RRNA Gene And Metagenomic Sequencing

BACKGROUND:Cervical cancer is the most common malignant tumor in the female reproductive tract,and its incidence is increasing year by year and becoming younger.HR-HPV of persistent infection is a major pathogenic fact or in cervical precancerous lesions and cervical cancer.Many studies have shown that cervico-vaginal microbiota plays an important role in HR-HPV infection and cervical lesions,but the specific mechanism remains unclear.Most of the previous research results in this field were based on 16S rRNA gene sequencing,but the technology only focused on the sequencing of marker sequences,resulting in incomplete gene information acquisition,and the accuracy of identification depth of bacteria could only reach the genus level.Metagenomic sequencing is a whole-genome sequencing of sample DNA,which can be accurate to the species level in the identification of bacteria.At the same time,it can analyze the function and metabolic pathway of microbiota,which can effectively make up for the deficiency of 16S rRNA gene sequencing.OBJECTIVE:Based on 16S rRNA gene and metagenomic sequencing,to analyze the composition,structure and function of the cervical microbiota of H R-HPV infection and normal women,and to explore the relationship between the cervical microbiota and HR-HPV infection,so as to provide clues for revea ling the mechanism of HR-HPV infection and cervical lesions.METHODS:From December 2020 to September 2021,40 women of child bearing age who met the inclusion criteria(20 women infected with HR-HPV and 20 normal women)in Guangdong Second Provincial General Hospital were selected as the sujects.Their cervical swabs were taken as samples for 16S rRNA and metagenomic sequencing,and bioinformatics analysis was performed.RESULTS:16S rRNA gene sequencing analysis results:1.The Alpha diversity indexes were significantly higher in the HR-HPV group than in the Control group.2.Beta diversity analysis suggested significant differences in the structure of cervical microbiota between the HR-HPV and Control groups.3.Compared with the Control group,the abundance of Firmicutes in the HR-HPV group was significantly decreased and Actinobacteria was significantly increased at the phylum level.At the genus level,the abundance of Lactobacillus decreased significantly in HR-HPV group,while the abundance of Gardner ella,Atopobium,and Bifidobacterium increased significantly.4.LEfSe analysis suggested that at the genus level,Lactobacillus,Gardner ella,Atopobium,and Bifidobacterium were the bacterias with biomarker significance among groups.Metagenomic sequencing analysis results:1.Analysis of bacterial composition and differences between groups at the phylum and genus level showed that the results of metagenomic sequencing were basically consistent with 16S rRNA gene sequencing.2.At the species level,compared with the Control group,the abundance of Lactobacillus crispatus was significantly decreased in the HR-HPV group,and the abundance of Gardnerella vaginalis was significantly increased.3.LEfSe analysis suggested that Lactobacillus crispatus,Lactobacillus jensenii,and Lactobacillus helveticus had high biomarker significance at the species level.4.HUMAnN2 functional analysis suggested that there were some metabolic pathways and metabolites associated with the abundence of cervical microbiota significantly different between the HR-HPV and Control groups.CONCLUSION:Cervical microbiota was associated with HR-HPV infection.The richness and diversity of cervical microbiota in women infected with HR-HPV were significantly increased,and the abundance of Lactobacillus crispat us,Lactobacillus jensenii,and Lactobacillus helveticus were significantly decreased,and the related metabolic pathways and metabolites may play a role in HR-HPV infection,which are expected to become clinical targets for eliminating HR-HPV infection and interfering with the progression of cervical lesions.