人源与牛源非O157产志贺毒素大肠杆菌耐药性分析及脉冲场凝胶电泳分子分型  

Drug resistance analysis and pulsed-field gel electrophoresis genotyping of non-O157 Shiga toxin-producing Escherichia coli isolated from human and cattle

作　　者：施旭辉 于婷婷 王芋丹 陈文霞 黄顺敏 王磊[1] 郑晓风[1] 谢金鑫 佟盼盼 SHI Xu-hui;YU Ting-ting;WANG Yu-dan;CHEN Wen-xia;HUANG Shun-min;WANG Lei;ZHENG Xiao-feng;XIE Jin-xin;TONG Pan-pan(College of Animal Medicine,Xinjiang Agricultural University,Urumqi 830052,China)

机构地区：[1]新疆农业大学动物医学学院,新疆乌鲁木齐830052

出　　处：《中国预防兽医学报》2022年第11期1224-1229,共6页Chinese Journal of Preventive Veterinary Medicine

基　　金：国家自然科学基金(31960695);自治区高层次人才引进工程项目(XJGCC2018080);新疆农业大学博士后科研流动站资助(214045);新疆农业大学自治区级大学生创新创业训练计划项目(S202110758027)。

摘　　要：为了解人源和牛源非O157产志贺毒素大肠杆菌(STEC)的耐药表型、耐药基因、质粒复制子类型及其之间的同源性,本研究采用K-B纸片法对3株人源非O157 STEC和14株牛源非O157 STEC耐药株进行药敏试验,结果显示3株人源非O157 STEC均耐药,其中2株呈多重耐药(MDR);14株牛源非O157 STEC耐药株中有9株呈MDR。11株MDR-非O157 STEC对4~13种抗生素耐药,包括β-内酰胺类、四环素类、磺胺类、氯霉素类和氨基糖苷类药物,其中10株(1株人源和9株牛源)非O157 STEC的耐药谱中出现氯霉素-链霉素-复方新诺明-四环素组合。通过PCR方法检测非O157 STEC耐药基因floR(氯霉素类)、mph(A)(大环内酯类)、strA、strB(氨基糖苷类)、sulR(磺胺类)和tet(A)、tet(G)(四环素类)和质粒复制子分型,结果显示17株非O157 STEC中有7株携带floR-mph(A)strA-strB-sulR-tet(A)-tet(G),2株携带floR-strA-strB-sulR-tet(A)-tet(G),2株携带tet(A)-tet(G),1株携带floR-mph(A)-tet(A)-tet(G),1株携带floR-mph(A),4株未携带检测的耐药基因;质粒复制子分型显示,IncFIB质粒检测率最高为82%(14/17),IncC和IncW质粒次之(47%,8/17)。通过脉冲场凝胶电泳(PFGE)对人源和牛源非O157 STEC耐药株进行同源性分析,结果显示,人源与牛源非O157 STEC耐药株的相似度低于80%。本研究上述结果表明人源和牛源非O157 STEC均出现了MDR,同源性不高,但这些菌株携带多个耐药基因和质粒,且呈现相似的耐药模式,并且本研究首次在我国新疆地区非O157 STEC中检测出mph(A)基因,提示在动物生产中需谨慎用药,以防止同时具有耐药性和毒力的菌株感染人类。In this study,the drug resistance phenotype,drug resistance genes,plasmid replicon type,and homology of Shiga toxin-producing Escherichia coli(STEC) from humans and cattle were analyzed.The Kirby-Bauer disk diffusion method was used to detect the drug sensitivity of three human non-O157 STEC isolates and 14 cattle non-O157 STEC isolates.The results showed that all three human non-O157 STEC isolates were drug-resistant.Among them,two were multi-drug resistant(MDR).Nine of 14 bovine STEC isolates were MDR.These MDR-non-O157 STEC isolates showed resistance to 4-13 antibiotics,includingβ-lactam,tetracycline,sulfonamides,amidoalcohols,and aminoglycosides.Ten isolates(one from humans and nine from cattle)showed resistance to the chloramphenicol,streptomycin,trimethoprim-sulfamethoxazole,and tetracycline combined.Drug resistance genes of floR(chloramphenicols),mph(A)(macrolides),strA,strB(aminoglycosides),sulR(sulfonamides),tet(A) and tet(G)(tetracyclines),and plasmid replicons type non-O157 STEC were tested by PCR.The results showed that seven isolates carried floR-mph(A)-strA-strB-sulR-tet(A)-tet(G),two isolates had floR-strA-strB-sulR-tet(A)-tet(G),two isolates carried tet(A)-tet(G),one isolate carried floR-mph(A)-tet(A)-tet(G),and one isolate carried floR-mph(A).In contrast,four isolates did not have the drug-resistance gene.Plasmid replicon typing revealed that the detection rate of IncFIB plasmid was the highest,with 82%(14/17),followed by IncC and IncW plasmids(47%,8/17).Homology analysis by pulsed-field gel electrophoresis(PFGE) showed that less than 80% homolog was observed between human and bovine non-O157 STEC-resistant strains.This study showed that both human and bovine non-O157 STEC isolates have MDR,while these MDR genes shared low homology.The mph(A) gene was first detected in non-O157 STEC isolates in Xinjiang,suggesting that it is necessary to use drugs carefully in animal production to prevent strains with both drug resistance and virulence from infecting humans.

关 键 词：产志贺毒素大肠杆菌 耐药性 质粒 腹泻患者 脉冲场凝胶电泳 

分 类 号：S852.61[农业科学—基础兽医学]