广泛耐药肺炎克雷伯菌耐药表型分析及耐药机制研究  

Drug resistance phenotype analysis and mechanism investigation of extensively drug resistant Klebsiella pneumoniae

作　　者：张天翼 周永年 栗子洋 戎建荣[1] Zhang Tianyi;Zhou Yongnian;Li Ziyang;Rong Jianrong(Department of Clinical Laboratory,Third Hospital of Shanxi Medical University,Shanxi Bethune Hospital,Shanxi Academy of Medical Sciences Tongji Shanxi Hospital,Taiyuan 030032,China)

机构地区：[1]山西医科大学第三医院山西白求恩医院山西医学科学院同济山西医院检验科,太原030032

出　　处：《中华检验医学杂志》2022年第9期936-942,共7页Chinese Journal of Laboratory Medicine

基　　金：山西省自然科学基金 (201901D111420)。

摘　　要：目的研究广泛耐药肺炎克雷伯菌(XDRKP)耐药表型特点及相关耐药机制。方法收集山西白求恩医院2018年1月至2020年12月内分离的3201株肺炎克雷伯菌并根据已有药敏结果筛选116株广泛耐药肺炎克雷伯菌纳入研究。基质辅助激光解吸电离飞行时间质谱(MALDI-TOF MS)及梅里埃VITEK-compact 2进行菌株鉴定及药敏试验,药敏结果K-B法或微量肉汤稀释法复核。改良碳青霉烯酶灭活试验(mCIM)联合乙二胺四乙酸协同碳青霉烯酶灭活试验(eCIM)检测XDRKP碳青霉烯酶表型,并与碳青霉烯酶基因扩增结果比对。聚合酶链反应(PCR)扩增耐药基因:碳青霉烯酶基因(blaKPC、blaNDM、blaVIM、blaIMP、blaOXA);氨基糖苷耐药基因:16S rRNA甲基化酶基因(rmtA、rmtC、rmtD、rmtG、rmtH、armA、npmA、rmtB、rmtE、rmtF),氨基聚糖苷修饰酶基因变体[aac(6′)-Ib-cr];喹诺酮耐药基因:DNA解旋酶保护蛋白qnr家族基因(qnrA、qnrB、qnrC、qnrD、qnrS),外排泵蛋白基因(oqxAB、qepA),氨基聚糖苷修饰酶基因变体[aac(6′)-Ib-cr];替加环素耐药Tet蛋白基因[外排泵蛋白基因:tet(A)和tet(L)],核糖体保护蛋白基因[tet(M)],替加环素修饰酶基因[tet(X)]。对照分析各XDRKP耐药表型与相应耐药基因携带情况间的关系。结果共收集到XDRKP 116株。头孢菌素类及喹诺酮类抗菌药物耐药率100%(116/116),碳青霉烯类抗菌药物耐药率99.14%(115/116),氨基糖苷类抗菌药物庆大霉素、妥布霉素、阿米卡星耐药率分别为95.69%(111/116)、94.83%(110/116)、88.79%(103/116),磺胺类抗菌药物耐药(44/116)与替加环素耐药(3/116)检出率较低。mCIM联合eCIM试验结果与碳青霉烯酶基因扩增结果一致率95.65%(110/115)。各耐药基因阳性率:blaKPC 90.52%(105/116),blaNDM 10.34%(12/116),rmtB 81.90%(95/116),armA 2.59%(3/116),oxqAB 65.52%(76/116),qnrB 6.03%(7/116)、qnrS 12.93%(15/116)、aac(6′)-Ib-cr 7.76%(9/116),tet(A)21.55%(25/116)。其余各耐药基因未检出。匹配分Objective This work aims to investigate the phenotype-characteristics of drug resistance and the possible mechanisms of extensively drug-resistance Klebsiella pneumoniae(XDRKP).Methods Screened by the previous drug susceptibility results,116 clinical Klebsiella pneumoniae isolates were collected from Shanxi Bethune Hospital from January 2018 to December 2020.Matrix-assisted laser desorption/ionization time of flight mass spectrometry(MALDI-TOF MS)rapid microbial identification system and VITEK-compact 2 were used.The modified carbapenem inactivation method(mCIM)combining with EDTA carbapenem inactivation method(eCIM)was used to identify the strains′carbapenemase phenotypes,which were compared with subsequent qPCR results.The qPCR amplification combining with agarose gel electrophoresis were carried out to detect various drug-resistant related genes,including:carbapenemase genes:blaKPC,blaNDM,blaVIM,blaIMP,blaOXA;aminoglycosides resistance genes:①16S rRNA methylase genes:rmtA,rmtC,rmtD,rmtG,rmtH,armA,npmA,rmtB,rmtE,rmtF,②variant of aminoglycosides acetyltransferase gene:aac(6′)-Ib-cr;quinolone resistance genes:DNA gyrase protection protein qnr family:qnrA,qnrB,qnrC,qnrD,qnrS,efflux pump protein gene:oqxAB,qepA,variant of aminoglycoside acetyltransferase gene:aac(6′)-Ib-cr;and tigecycline-resistant Tet protein genes:efflux pump protein gene:tet(A),tet(L),ribosome protection protein gene:tet(M),tigecycline modified enzyme gene:tet(X).Each isolate′s phenotype and resistance gene result were compared and analyzed correspondingly.Results A total number of 116 XDRKP isolates were collected in 3 years,115 of which are identified as carbapenem resistant.Both cephalosporins and quinolones resistant rate were 100%,while the resistant rate of aminoglycosides antibiotic gentamicin,tobramycin and amikacin was 95.69%(111/116),94.83%(110/116),or 88.79%(103/116)respectively.Sulfonamide antibiotics and tigecycline showed a relatively lower resistant rate.Compared with PCR amplification results,mCIM combining with eCIM

关 键 词：克雷伯菌 肺炎 广泛耐药 卡巴配能类 耐药机制 

分 类 号：R446.5[医药卫生—诊断学]