Correlation Between HPV Infection And Cervical Microbiota Among Women In Inner Mongolia

Objective:The aim of this research is to ascertain the prevalence and type of HPV in women of Inner Mongolia,thereby furnishing an epidemiological foundation for HPV inoculation,prevention,treatment,and monitoring of cervical lesions in the area.At the same time,by evaluating the detection efficiency of different kits,it is intended to screen out more efficient kits,and to improve the screening rate of the virus to a certain extent.This study further analyzed the diversity of cervical microflora under HPV infection in Inner Mongolia,explored the relationship between HPV infection and cervical microbiota,and predicted the function of cervical microbiota,in order to provide new strategies for the prevention,control and treatment of HPV.Method:Firstly,4068 cases of HPV genotyping test results were collected from the Inner Mongolia Peoples Hospital.SPSS was used for statistical analysis to understand the infection rate,type distribution,infection mode and age composition of infected women in Inner Mongolia.Then,72 samples were tested for HPV typing by PCR-capillary electrophoresis fragment analysis and PCR-reverse dot hybridization.The results were compared,and the samples with inconsistent results were verified by generation sequencing.Finally,48 samples with single HPV infection,35 samples with multiple HPV infection and 30 samples without HPV infection were collected in parallel after the above two methods were used for HPV typing detection of cervical brush samples.The collected HPV-positive samples were set as the experimental group(divided into single infection group and multiple infection group),and the HPV-negative samples were set as the control group.After 16S r DNA sequencing,the Alpha diversity,Beta diversity and community composition structure of different groups were studied by bioinformatics analysis and the different bacteria among the groups were found,the function of cervical microbiota were predicted,and explored the relationship between HPV infection and cervical microbiota.Results:HPV infection in Inner Mongolia:the total infection rate of HPV was 20.26%,14.87%of the people were infected with single genotype,5.38%of the people were infected with multiple genotype,and at most six subtypes of simultaneous infection were detected.The two most common types are HPV52 and HPV16,while the most common genotype combination is HPV66+52.There were significant differences in the infection rate among different age groups(X~2=18.892,P<0.01),and the infection rate was highest in the age group of sixty years and above.The results of 4 samples were found inconsistent by the two methods,and the total coincidence rate was 94.44%(Kappa=0.846).After the sanger sequencing verification,it was found that the sequencing results of other samples were consistent with the results of PCR-capillary electrophoresis fragment analysis,except for sample No.72.16S r DNA sequencing analysis showed that alpha diversity in single infection group and multiple infection group was significantly higher than that in control group(P<0.05).Beta diversity was significantly different among three groups(P<0.05).The composition of cervical microbiota was different among the three groups:at the phylum level,the proportion of Firmicutes in the control group was highest,while the proportion of Firmicutes in the single infection group and the multiple infection group was decreased,and the proportion of Actinobacteriota,Proteobacteria and Bacteroidota was increased.At the genus level,the relative abundance of Lactobacillus was decreased in the single infection group and the multiple infection group,while the relative abundance of Gardnerella,Prevotella and Atopobium were increased.LEf Se was used to further analyze the species with significant differences among the three groups.A total of 22 bacterial branches showed significant differences in relative abundance among the single infection,the multiple infection and the control group.The control group was significantly enriched in Lactobacillus,while the multiple infection group was significantly enriched in Bacteroidetes,Prevotella and Veronococcus.The random forest model was constructed using the different bacteria genus,and the AUC value was 0.71,indicating that the different bacteria community could predict HPV infection to a certain extent.By functional prediction analysis,the metabolic pathways,biosynthesis of secondary metabolites,biosynthesis of amino acids,carbon metabolism,purine metabolism,cysteine and methionine metabolism,alanine-aspartate and glutamate metabolism of HPV infection group were higher than those of control group(P<0.05).The metabolic activity of the cervical microbiota may provide a favorable microenvironment for the survival of HPV and other pathogens.The phosphotransferase system,mismatch repair and galactose metabolism in control group were significantly higher than those in HPV infection group(P<0.05).These metabolic pathways may be one of the mechanisms against bacterial dysregulation and HPV infection.Among the eight microbial phenotypes,the relative abundance of gram-negative bacteria was higher in the single infection group and the multiple infection group(P<0.05),and the relative abundance of Gram-positive bacteria and mobile elements was higher in the control group(P<0.05).In addition,the relative abundance of anaerobic and biofilm-forming phenotypes was higher in the multiple infection group than in the control group(P<0.01).Conclusion:There is a correlation between cervical microbiota and HPV infection.The species diversity of cervical microbiota in both single and multiple infection groups was higher than that in control group.Lactobacillus is the protective factor of HPV infection,Prevotella,Gardnerella and other anaerobic bacteria are closely related to HPV infection,which is of great significance for predicting HPV infection.In addition to the HPV infection-related microbiota reported so far,this study especially found that Veillonellaceae may be associated with HPV infection,which provided new ideas for the development of strategies to prevent and control HPV infection-related diseases from the perspective of maintaining the balance of cervical microbiota.