质粒介导的印第安纳沙门氏菌耐药机制及分子溯源研究  

Plasmid-mediated mechanism and traceability of pan-resistance in Salmonella indiana

作　　者：王天 徐红红 张林吉[2] 王学锋 张小荣[3] 吴艳涛[3] 巢国祥 WANG Tian;XU Hong-hong;ZHANG Lin-ji;WANG Xue-feng;ZHANG Xiao-rong;WU Yan-tao;CHAO Guo-xiang(School of Public Health Nursing,Yangzhou University,Yangzhou 225001,China;Xuzhou Polytechnic College of Bioengineering,Xuzhou 221006,China;College of Veterinary Medicine,Yangzhou University,Yangzhou 225009,China;Yangzhou Center for Disease Control and Prevention,Yangzhou 225002,China)

机构地区：[1]扬州大学护理学院公共卫生学院,扬州225001 [2]徐州生物工程职业技术学院,徐州221006 [3]扬州大学兽医学院,扬州225009 [4]扬州市疾病预防控制中心,扬州225002

出　　处：《中国人兽共患病学报》2022年第7期619-625,共7页Chinese Journal of Zoonoses

基　　金：科技部科技助力经济2020重点专项项目(No.SQ2020YFF0426460);国家重点研发计划项目(No.2016YFD0501600);徐州市科技项目(No.KC21297)。

摘　　要：目的通过全基因测序方法(NGS)分析印第安纳沙门氏菌耐药克隆的质粒结构特征并与肠杆菌科质粒BLAST分子溯源,通过质粒消除研究其耐药表型和基因型的变化,探索印第安纳沙门氏菌耐药克隆成因及起源。方法通过对2株(编号222、15)泛耐药印第安纳沙门菌全基因测序结果尤其是其质粒序列耐药I类整合子、耐药基因、移动元件的分析,并与NCBI中28株肠杆菌科菌的质粒BLAST比对溯源。对3株印第安纳沙门氏菌(编号15、38、222),1株肠炎沙门氏菌(编号17)、1株鼠伤寒沙门氏菌(编号32)、1株德尔卑沙门氏菌(编号163)进行高温-SDS质粒消除试验并比较质粒消除后菌株耐药表型及基因型的变化。结果全基因测序结果显示5株菌株拥有1个带有1个以上的耐药I类整合子及多个转座酶(Tns)和插入序列(ISs)等多种移动元件的结构相似、携带大量耐药基因且同源性大于99%的质粒(>210000 bp)。经质粒消除实验后菌株15、38对四环素、庆大霉素、环丙沙星、阿奇霉素、阿米卡星等抗生素恢复敏感,菌株222对头孢噻圬、庆大霉素、环丙沙星、萘啶酸、卡那霉素等抗生素恢复敏感。菌株15不再携带TEM-1、OXA-1、sul1、aacC4、dfrA17、floR、qnrB等7种耐药基因,菌株38不再携带TEM-1、OXA-1、CTX-M、sul1、aacC4、dfrA17、floR,菌株222不再携带TEM-1、OXA-1、sul1、aacC4、aac6-1b。BLAST溯源结果表明印第安纳携带的质粒与大肠杆菌质粒高度相似,同源性大于99%。结论印第安纳沙门氏菌拥有含耐药I类整合子、多种移动元件(Tns)、插入序列(ISs)及大量耐药基因的耐药质粒是印第安纳沙门氏菌泛耐药克隆形成的主要原因。该质粒可能是其进化过程中从大肠杆菌类菌中获得。Next generation sequencing(NGS)was used to perform molecular characterization of the resistance plasmids of Salmonella indiana clones,and their origins were traced with a BLAST search against Enterobacteriaceae plasmids.Plasmid elimination tests were performed to study the variations in pheno types and genotypes of S.indiana to further confirm the roles of the resistance plasmids.The plasmid NGS results of two S.indiana strains(No.222,15)were analyzed on the basis of class I integrons,resistance genes and mobile elements,and their origins were traced with a BLAST search against the other 28 plasmids of Enterobacteriaceae in NCBI.Plasmid elimination and resistance tests were performed on three S.indiana(No.15,38,222),one S.enterica(No.17),one S.typhimurium(No.17)and one S.derby(No.163)isolates.The results of NGS showed that all five S.indiana strains had a structurally similar large plasmid(>210000 bp)with class I integron(s),many mobile elements,ISs and resistance genes;the sequence identity was>99%.The results of plasmid elimination and resistance tests showed that the phenotypes and genotypes of S.indiana had corresponding varies.The BLAST tracing results indicated that the plasmid(s)were highly similar to E.coli plasmid(s)(identity>99%).S.indiana harbored plasmid(s)with class I integron(s),many different mobile elements,such as Tns,ISs and resistance genes,thus indicating the formation mechanism of the S.indiana pan-resistant clone.

关 键 词：印第安纳沙门氏菌 全基因组测序(NGS) 耐药质粒 质粒消除 

分 类 号：R378.2[医药卫生—病原生物学]