肝内胆管结石中抗生素耐药基因在性别之间分布差异的初步分析

A preliminary analysis of the sex distribution of antibiotic resistance genes in patients with intrahepatic bile duct stones

目的分析肝内胆管结石抗生素耐药基因(ARG)在不同性别之间的分布,为临床合理使用抗生素提供参考。方法收集2017年10月-2018年9月山西医科大学附属人民医院收治的肝内胆总管结石患者共120例,其中男60例,女60例,对所采集标本进行细菌基因组提取,并采用实时定量PCR方法进行检测。符合正态分布的计量资料组间比较采用t检验;不符合正态分布的计量资料组间比较采用Mann-Whitney U检验。Pearson线性相关分析确定可移动基因元件(MGE)和ARG丰度之间是否具有显著相关性。结果在所有测得的ARG中,男性ARG 16种,其中sull的基因检出率最高(9. 4%)。blaCTX-M、blaTEM、blaSHV、blaampC、ampC-01、ampC/blaDHA、bla-ACC-1、blaCMY、blaGES、cfxA、aac、aacc、aadA1、strB、tetX的检出率分别为7. 6%、6. 9%、7. 3%、7. 8%、6. 2%、6. 5%、3. 9%、5. 5%、2. 4%、3. 9%、5. 6%、7. 4%、6. 8%、4. 2%、8. 6%。女性ARG 7种,其中sull的基因检出率最高(24%)。blaTEM、aacc、aadD、aadA1、strB、tetX的检出率分别为11. 3%、10. 8%、8. 4%、11. 9%、23. 9%、9. 8%。aadD在男性中未被检出; blaCTX-M、blaSHV、blaampC、ampC-01、ampC/blaDHA、bla-ACC-1、blaCMY、blaGES、cfxA、aac在女性中未被检出。16S rRNA基因与β-内酰胺类ARG、四环素类ARG具有显著相关性(P <0. 01),与氨基糖苷类ARG、磺胺类ARG具有相关性(P <0. 05); tnpA基因与β-内酰胺类ARG、四环素类ARGs和磺胺类ARG具有相关性(P <0. 05),与intⅠ基因具有显著相关性(P <0. 01)。结论肝内胆管结石中携带非常丰富的ARG,其中sull的基因丰度最高;同时,男性中的ARG丰度高于女性,女性中β-内酰胺类ARG的丰度较少。因此,对于肝内胆管结石患者进行抗感染治疗时,女性应考虑将β-内酰胺类抗生素作为首选,而男性患者应给予更多剂量的抗生素或考虑多联用药。

Objective To investigate the distribution of antibiotic resistance genes (ARGs) in patients with intrahepatic bile duct stones,and to provide a theoretical basis for rational use of antibiotics in clinical practice. Methods A total of 120 patients with intrahepatic bile duct stones (60 male and 60 female patients) in People′s Hospital Affrliated to Shanxi Medical Universiry,from October 2017 to Septemper 2018 were enrolled. Bacterial genome was extracted from the collected specimens,and PCR was used for detection. The t -test was used for comparison of normally distributed continuous data between groups,and the Mann-Whitney U test was used for comparison of non-normally distributed continuous data between groups. Pearson correlation analysis was used to investigate the correlation of normally distributed continuous data. Results Off all ARGs detected,16 were detected in male patients,among which the sull gene had the highest detection rate (9.4%);in male patients,the detection rates of blaCTX-M,blaTEM,blaSHV,blaampC,ampC-01,ampC/blaDHA,bla-ACC-1,blaCMY,blaGES,cfxA,aac,aacc,aadA1,strB,and tetX were 7.6%,6.9%,7.3%,7.8%,6.2%,6.5%,3.9%,5.5%,2.4%,3.9%,5.6%,7.4%,6.8%,4.2%,and 8.6%,respectively. Off all ARGs detected,7 were detected in female patients,among which the sull gene had the highest detection rate (24%);in female patients,the detection rates of blaTEM,aacc,aadD,aadA1,strB,and tetX were 11.3%,10.8%,8.4%,11.9%,23.9%,and 9.8%,respectively. The aadD gene was not tested in male patients,and blaCTX-M,blaSHV,blaampC,ampC-01,ampC/blaDHA,bla-ACC-1,blaCMY,blaGES,cfxA,and aac were not detected in female patients. 16S rRNA gene was significantly correlated with beta-lactam ARG and tetracycline ARG( P <0.01),aminoglycoside ARG and sulfonamide ARG( P <0.05),tnpA gene was significantly correlated with beta-lactam ARG,tetracycline ARGs and sulfonamide ARG( P <0.05),and intI gene( P <0.01). Conclusion There are abundant antibiotic resistance genes in patients with intrahepatic bile duct stones,among which the sull gene has the highest abundance. At the same time,male patients have higher abundance of genes than female patients,and female patients have low abundance of beta-lactam ARGs. Therefore,beta-lactam antibiotics should be the first choice for anti-infective therapy for female patients with intrahepatic bile duct stones,while men should use a higher dose of antibiotics or a combination of several antibiotics.