肛肠手术切口感染病原菌对喹诺酮类药物耐药性及机制分析  

Analysis of quinolones in the treatment of pathogens causing an anorectal surgical site infection

作　　者：路平[1] 李瑶[2] 南颖 LU Ping;LI Yao;NAN Ying(Beijing Rectum Hospitals Beijing Er Long Lu Hospital,Beijing,China 100120;The Third Hospital of the Chaoyang District,Beijing;Beijing Fengtai You An Men Community Health Service Center,Fengtai District)

机构地区：[1]北京市肛肠医院(北京市二龙路医院),北京西城100120 [2]北京市朝阳区第三医院 [3]北京市丰台区右安门社区卫生服务中心

出　　处：《中国病原生物学杂志》2019年第9期1076-1080,共5页Journal of Pathogen Biology

摘　　要：目的研究喹诺酮类药物对肛肠手术切口病原菌感染治疗效果及相关机制,为临床用药提供依据。方法选取2016年1月-2017年12月北京地区316例肛肠手术患者临床资料,无菌采集患者手术切口处脓液或分泌物,全自动微生物鉴定系统进行菌株鉴定,K-B纸片法测定受试菌株对萘啶酸、诺氟沙星,左氧氟沙星,环丙沙星和加替沙星敏感性。采用微量稀释法测定大肠埃希菌对萘啶酸、诺氟沙星,左氧氟沙星,环丙沙星和加替沙星敏感性。利用PCR方法检测大肠埃希菌gyrA、gyrB、parC、parE、OmpC和OmpF基因并进行测序。结果316份标本中共有52份标本病原菌培养呈阳性,阳性率16.46%。分离出52株病原菌中革兰阴性菌46株,革兰阳性菌5株,真菌1株。革兰阴性菌中大肠埃希菌35株,变形菌属5株,克雷伯菌属4株,肠杆菌属2株;革兰阳性菌中葡萄菌属3株,肠球菌属2株;真菌为假丝酵母菌。经ESBLs确证试验共筛选出产ESBLs大肠埃希菌16株,检出率为45.71%。革兰阴性菌分离株对萘啶酸、诺氟沙星、左氧氟沙星、环丙沙星和加替沙星耐药菌株为:34、25、18、17和9株,耐药率分别为73.91%、54.35%、39.13%、36.96%和19.57%。革兰阳性菌分离株仅对萘啶酸和诺氟沙星耐药,分别为3株和1株。成功扩增出gyrA、gyrB、parC、parE、OmpC和OmpF基因。gyrA发生错义突变分别为Ser83→Leu、Ala84→Pro、Asp87→Asn和Asp87→Gly;gyrB发生错义突变分别为Phe361→Try、Glu470→Asp、Ser492→Asn;parC发生错义突变分别为Ser80→Ile、Glu84→Lys;parE发生错义突变分别为Asp420→Asn、Ala425→Val、Ser458→Ala;OmpC发生错义突变分别为Asp18→Glu,Val29→Lys和Ser64→Glu。讨论大肠埃希菌是肛肠手术切口感染的主要病原菌,大肠埃希菌喹诺酮类耐药决定区基因突变是对喹诺酮类抗生素产生耐药的主要原因,孔蛋白OmpC基因突变会导致大肠埃希菌对药物敏感性下降。Objective To study the therapeutic effect of quinolones on pathogens causing an infection after anorectal surgery and their related mechanism in order to provide a basis for clinical use of medication.Methods Clinical data on 316 patients undergoing anorectal surgery in Beijing from January 2016 to December 2017 were selected.Pus or secretions from the surgical site were collected aseptically,and the bacteria were identified by an automated microbial identification system.The susceptibility of the tested strains to naphthoic acid,norfloxacin,levofloxacin,ciprofloxacin,and gatifloxacin was determined using the K-B disc method.The sensitivity of Escherichia coli to nalidixic acid,norfloxacin,levofloxacin,ciprofloxacin,and gatifloxacin was determined using microdilution.The gyrA,gyrB,parC,parE,OmpC,and OmpF genes of E.coli were detected using PCR and sequenced.Results Of 316 specimens,52(16.46%)were positive according to a pathogen culture.The 52 strains of pathogens included 46 strains of Gram-negative bacteria,5 strains of Gram-positive bacteria,and 1 strain of fungus.Gram-negative bacteria included 35 strains of E.coli,5 strains of Proteus,4 strains of Klebsiella,and 2 strains of Enterobacter.Gram-positive bacteria included 3 strains of Staphylococcus and 2 strains of Enterococcus.The fungus was Candida.Screening for ESBL production revealed that 16 strains(45.71%)of E.coli produced ESBLs.Thirty-four strains of Gram-negative bacteria were resistant to nalidixic acid,25 were resistant to norfloxacin,18 were resistant to levofloxacin,17 were resistant to ciprofloxacin,and 9 were resistant to gatifloxacin.Three strains of Gram-positive bacteria were resistant to nalidixic acid and 1 was resistant to norfloxacin.The gyrA,gyrB,parC,parE,OmpC,and OmpF genes were successfully amplified.The missense mutations in gyrA were Ser83→Leu,Ala84→Pro,and Asp87→Asn.The missense mutations in gyrB were Phe361→Try,Glu470→Asp,and Ser492→Asn.The missense mutations in parC were Ser80→Ile and Glu84→Lys.The missense mutati

关 键 词：喹诺酮类药物 肛肠手术切口 病原菌 大肠埃希菌 

分 类 号：R378[医药卫生—病原生物学]