关于胆囊结石、胆囊结石合并胆总管结石、胆总管结石抗生素抗性基因的初步研究

Preliminary study on antibiotic-resistance genes of bacteria in patients with gallbladder calculi, gallbladder calculi combined with common bile duct calculi, and common bile duct calculi

目的了解抗生素抗性基因在胆石病患者中的分布状况,为临床合理使用抗生素提供理论依据。方法收集胆囊结石30例、胆囊结石合并胆总管结石30例、胆总管结石30例,对采集到的标本进行细菌基因组的提取,并采用qPCR的方法对提取的基因组进行检测。结果在检测到的ARGs中,胆囊结石患者β-内酰胺类抗生素抗性基因占的比例最低为5.92%,检测到的基因有8种,strB基因检出率最高,为27.1%。blaTEM、cfxA、aacc、aadD、aadA1、tetX和sull的检出率分别为13.9%、5.1%、5.1%、7.7%、20.7%、4.6%和15.6%。胆囊结石合并胆总管结石患者β-内酰胺类抗生素抗性基因占的比例最低为12.94%,检测到的基因有15种,sull基因检出率最高,为17.1%。blaCTX-M、blaTEM、blaSHV、ampC-01、ampC/blaDHA、blaACC-1、blaGES、cfxA、aac、aacc、aadA1、strB和tetX的检出率分别为3.2%、9.4%、4.4%、3.4%、8.9%、4.2%、2.8%、2.5%、8.2%、3.9%、8.1%、6.2%、8.2%和9.4%。胆总管结石患者氨基糖苷类抗生素抗性基因占的比例最低为17.34%,检测到的基因有16种,blaTEM基因检出率最高,为9.1%。blaCTX-M、blaSHV、ampC-01、ampC/blaDHA、bla-ACC-1、blaCMY、blaGES、cfxA、aac、aacc、aadA1、str B、tet X和sull的检出率分别为7.3%、6.5%、7.6%、5.4%、5.6%、3.3%、6.4%、4.0%、4.5%、4.7%、6.9%、6.1%、6.0%、7.8%和8.9%。blaCMY只在胆总管结石被测到,aad D只在胆囊结石中被测到。结论ARGs不仅广泛存在于公共环境中,也存在于人体胆道系统中;同时,在胆道系统中,胆总管结石携带ARGs最多,胆囊结石合并胆总管结石次之,胆囊结石携带的ARGs最少;胆囊结石患者和胆囊结石合并胆总管结石患者中β-内酰胺类抗生素抗性基因含量最低;胆总管结石患者中氨基糖苷类抗生素抗性基因含量最低。这些发现可以为胆石病患者在抗生素的选择上提供理论依据。

Objective To understand the distribution of antibiotic resistance genes in patients with cholelithiasis and provide theoretical basis for rational use of antibiotics in clinic.Methods 30 cases of gallbladder stones,30 cases of gallbladder stones combined with choledocholithiasis and 30 cases of choledocholithiasis were collected.The bacterial genomes were extracted from collected specimens,and the extracted genomes were detected by PCR.Results Among all detected ARGs,5.92%of the patients with gallbladder stones had the lowest proportion of bacteria with antibiotic-resistance genes to beta-lactam.There were eight antibiotic-resistance genes of bacteria in patients with gallstone stone,and the detection rate of str B gene was the highest,27.1%.The detection rates of blaTEM,cfx A,aacc,aad D,aad A1,tetX and sull were 13.9%,5.1%,5.1%,7.7%,20.7%,4.6%and 15.6%respectively.Bacteria in 12.94%of the patients with gallbladder stones complicated with choledocholithiasis had the lowest proportion ofantibiotic-resistance genes to beta-lactam.There were 15 antibiotic-resistance genes of bacteria in patients withgallstone complicated with choledocholithiasis,and the detection rate of sull gene was the highest,17.1%.The detectionrates of blaCTX-M,blaTEM,blaSHV,ampC-01,amp C/blaDHA,bla-ACC-1,blaGES,cfx A,aac,aacc,aad A1,str B,and tetX were 3.2%,9.4%,4.4%,3.4%,8.9%,4.2%,2.8%,2.5%,8.2%,3.9%,8.1%,6.2%,8.2%,8.2%,and 9.4%respectively.The lowest proportion of aminoglycoside antibiotic resistance genes in patients with choledocholithiasis was 17.34%.There were 16 types of choledocholithiasis genes,with the highest detection rate of blaTEM gene,9.1%.The detection rates of blaCTX-M,blaSHV,amp C-01,amp C/blaDHA,bla-ACC-1,blaCMY,blaGES,cfx A,aac,aacc,aad A1,strB,tet X,and sull were 7.3%,6.5%,7.6%,5.4%,5.6%,4.0%,4.0%,4.5%,4.7%,6.9%,6.9%,6.1%,6.0%,7.8%and 8.9%,respectively.The gene blaCMY was only detected in patients with choledocholithiasis,and aad D was only detected inpatients with gallbladder stone.Conclusion Antibiotic-resistance genes(ARGs)exist not only in the public environment,but also in the human biliary tract system.At the same time,in the biliary tract system,common bile duct stones carry the most ARGs,followed by common bile duct stones combined with gallbladder stones,and gallbladder stones carry the least ARGs.It was found that the lowest level of beta-lactam antibiotic-resistance genes was found in patients with gallbladder stones and common bile duct stones combined with gallbladder stones,and the lowest level of aminoglycoside antibiotic-resistance genes was found in patients with common bile duct stones.These findings can provide a theoretical basis for the selection of antibiotics in patients with cholelithiasis.