Analysis Of Bile Microorganism Difference Between Cholangiocarcinoma And Biliary Tract Benign Lesions

Objective:16S rRNA high-throughput sequencing was used to analyze the composition and difference of biliary tract flora in patients with cholangiocarcinoma and common bile duct stones.Methods:Prospectively enrolled patients with cholangiocarcinoma and Common bile duct stones,collected the clinical data of the patients,and collected the bile samples of the patients’common bile duct during ERCP and surgical operations,and performed DNA extraction and 16S rRNA sequencing.Analyze the composition and differences of bile microbes between patients with cholangiocarcinoma and patients with common bile duct stones,and explore the correlation between bile microbes and cholangiocarcinoma.Results:The study finally included 15 patients with cholangiocarcinoma(tumor group)and 20 patients with common bile duct stones(control group).The detection rate of microorganisms in all samples is 100%.The most common phyla in cholangiocarcinoma bile samples are Actinobacteria(63.39%),Proteobacteria(29.99%),Bacteroides(3.74%)and Firmicutes(2.66%);the most common families are Micrococcaceae(63.28%),Enterobacteriaceae(16.83%),Xanthomonasae(7.47%)and Pseudomonas(3.08%);There are 284 genera in patients with cholangiocarcinoma and bile duct benign diseases,among which the most common genera in bile samples from cholangiocarcinoma are Escherichia(58.29%),Paenarthrobacter(9.81%),and Stenotrophomonas(7.46%),Klebsiella(6.34%)and Pseudarthrobacter(4.20%),revealing the heterogeneity between individuals.Analysis of differences between Alpha diversity groups showed that there was no significant difference in bile microbiota composition between the tumor group and the control group(Chao1:P=0.76,Shannon:P=0.42),which did not have clinical significance;Beta diversity analysis The P values of T-test based on Weighted Unifrac and Unweighted Unifrac were 0.184 and 0.700,respectively,suggesting that the difference between the two groups of samples is not significant.However,the analysis of species differences showed that the characteristic flora of patients with cholangiocarcinoma were:Myroides,Flavobacteriales,Flavobacteriaceae,Singulisphaera,Klebsiella,Enterobacteriaceae,Enterobacterales,Acinetobacter and Moraxellaceae.Conclusion:Patients with cholangiocarcinoma have biliary tract microbiota imbalance,and changes in the composition of the biliary flora have potential biological and clinical significance for the occurrence and development of cholangiocarcinoma.