基于综合抗性基因数据库研究介导幽门螺杆菌对克拉霉素和左氧氟沙星耐药的易感基因  

Study on the susceptibility genes that mediate the resistance of Helicobacter pylori to clarithromycin and levofloxacin based on comprehensive antibiotic research database comparison

作　　者：吕涛 倪丽 陈川 孙洋洋 赵培微 吴娟 张嘉盛 司徒伟基 Lyu Tao;Ni Li;Chen Chuan;Sun Yangyang;Zhao Peiwei;Wu Juan;Cheung Kashing;Seto Waikay(Department of Gastroenterology and Hepatology,the University of Hong Kong-Shenzhen Hospital,Shenzhen 518053,China)

机构地区：[1]香港大学深圳医院消化及肝脏科,深圳518053

出　　处：《中华消化杂志》2022年第2期103-110,共8页Chinese Journal of Digestion

基　　金：广东省医学科学技术研究基金项目(B2021239);香港大学深圳医院医学研究重点项目(HKUSZH201902019)。

摘　　要：目的以拥有不同抗菌药物耐药性的幽门螺杆菌(H.pylori)临床菌株为研究对象,基于全基因组测序探索H.pylori对克拉霉素和左氧氟沙星(LVX)耐药相关新基因。方法纳入2016年9月1日至2019年8月31日在香港大学深圳医院消化及肝脏科因上消化道相关症状就诊且13C尿素呼气试验阳性的1749例患者,在行胃黏膜活体组织检查后进行胃黏膜组织H.pylori分离培养,成功保存H.pylori菌株90株。依据体外药敏试验结果,分别筛选克拉霉素单耐药(克拉霉素组)10株,LVX单耐药(LVX组)10株,克拉霉素和LVX双重耐药(双重耐药组)10株,克拉霉素、LVX、阿莫西林、呋喃唑酮、四环素、甲硝唑全敏感(全敏感组)10株,总计40株H.pylori菌株。通过全基因组测序,与综合抗性基因数据库(CARD)进行比对,分析单核苷酸变异(SNV)和插入缺失突变情况,筛选H.pylori对克拉霉素和LVX耐药相关基因并分析耐药相关新基因在4组间的表达差异。统计学方法采用独立样本t检验、单因素方差分析、最小显著差数法和卡方检验。结果克拉霉素组、LVX组、双重耐药组、全敏感组SNV位点数[(74952.00±8755.21)、(77128.10±3191.35)、(78639.90±601.23)、(77474.60±2421.05)个]和插入缺失突变数[(2582.20±265.45)、(2653.60±108.37)、(2667.10±43.82)、(2641.10±80.25)个]比较差异均无统计学意义(均P>0.05)。与CARD进行比对,共检出223个耐药相关基因,其中克拉霉素组克拉霉素单耐药相关基因19个,LVX组LVX单耐药相关基因24个,双重耐药组中有克拉霉素单耐药相关基因16个,LVX单耐药相关基因14个,双重耐药相关基因12个;全敏感组中有克拉霉素单耐药相关基因11个,LVX单耐药相关基因17个,双重耐药相关基因13个。克拉霉素单耐药相关基因中,红霉素酯酶基因(ere)B在克拉霉素组、LVX组、双重耐药组、全敏感组的检出率(0/10、0/10、3/10、0/10)比较差异有统计学意义(χ^(2)=5.79,P=0.049);红�Objective Taking clinical strains of Helicobacter pylori(H.pylori)with different antimicrobial resistance as the research object,to explore the new genes related to the resistance of H.pylori to clarithromycin(CLA)and levofloxacin(LVX)based on whole-genome sequencing.Methods From September 1st,2016 to August 31st,2019,1749 patients with upper gastrointestinal symptoms and positive 13C urea breath test who visited the Department of Gastroenterology and Hepatology,the University of Hong Kong-Shenzhen Hospital were enrolled.After gastric mucosal biopsy,H.pylori was isolated and cultured from gastric mucosa.Ninety H.pylori strains were successfully preserved.According to the results of in vitro drug sensitivity test,a total of 40 strains including 10 strains with single-drug resistance to CLA(CLA group),10 strains with single-drug resistance to LVX(LVX group),10 strains with dual-resistance to CLA and LVX(dual resistance group)and 10 strains sensitive to CLA,LVX,amoxicillin,furazolidone,tetracycline and metronidazole(all sensitive group)were screened out.By whole-genome sequencing and comparison to the comprehensive antibiotic research database(CARD),single nucleotide variations(SNV)and indels were analyzed,genes related to H.pylori resistance to CLA and LVX were screened out and the differences of new genes among 4 groups were analyzed.Independent sample t test,one-way analysis of variance,least significant difference method and chi-square test were used for statistical analysis.Results Among the 4 groups there were no statistically significant differences in the number of SNV(74952.00±8755.21,77128.10±3191.35,78639.90±601.23 and 77474.60±2421.05)and Indels(2582.20±265.45,2653.60±108.37,2667.10±43.82 and 2641.10±80.25)(all P>0.05).Compared to CARD,a total of 223 drug resistance-related genes were detected,of which 19 genes related to CLA mono-resistance in CLA group,24 genes related to LVX mono-resistance in LVX group,16 genes related to CLA mono-resistance,14 genes related to LVX mono-resistance,and 12 dual

关 键 词：螺杆菌 幽门 克拉霉素 左氧氟沙星 msrC qnr家族 外排转运系统 

分 类 号：R446.5[医药卫生—诊断学]