多药耐药鲍曼不动杆菌22种β-内酰胺酶基因和酶活性的检测研究  

Detection of 22 beta-lactamase genes and enzyme activities in multi-drug resistant Acinetobacter baumannii

作　　者：朱健铭 吴晋兰[3] 姜如金[1] 吴康乐[1] 王建敏[2] 孔海深[4] 

机构地区：[1]杭州市余杭区中医院检验科,311106 [2]杭州市余杭区中医院药剂科,311106 [3]杭州市余杭区第五人民医院内科 [4]浙江大学医学院附属第一医院检验科

出　　处：《中华临床感染病杂志》2009年第5期281-287,共7页Chinese Journal of Clinical Infectious Diseases

基　　金：基金项目：杭州市科技发展计划项目（20090833835）;杭州市卫生科技计划项目（2008e,0113）;杭州市余杭区重大科技项目（2008001）

摘　　要：目的了解临床分离的多药耐药鲍曼不动杆菌（multi—drug resistant Acirtetobacter baumannii，MDRAB）3-内酰胺酶基因存在状况以及酶活性。方法从浙江大学医学院附属第一医院住院患者中分离62株MDRAB，采用PCR及序列分析方法分析22种β-内酰胺酶基因，同时采用酶提取物改良三维试验检测超广谱β-内酰胺酶（ESBLs）、头孢菌素酶（AmpC酶）和金属β-内酰胺酶活性。结果62株MDRAB中，TEM、OXA-23群和ADC基因阳性株数分别为51株（82．3％）、50株（80．6％）和36株（58．1％），其余19种基因均为阴性；任选5株TEM基因PCR阳性产物测序比对，显示与GenBank基因库中TEM-1型相同；任选6株OXA-23群基因PCR阳性产物测序比对，显示与OXA-23型碳青霉烯酶基因（登录号：CAB69042．1）相同；任选2株ADC基因测序比对，显示与ADC-like型AmpC酶基因（登录号：EU081908）相同。有32株（51．6％）同时产ESBLs和AmpC酶，19株（30．6％）单独产ESBLs，1株（1．6％）单独产AmpC酶；金属β-内酰胺酶活性检测均为阴性。结论产OXA-23群碳青霉烯酶和ADC型AmpC酶是本组MDRAB对β-内酰胺类药物耐药的主要原因。Objective To investigate the distribution of 22 beta-laetamase genes and enzyme activities in multi-drug resistant Aeinetobacter baurnannii （MDRAB）. Methods Sixty-two MDRAB strains were isolated from the First Affiliated Hospital, College of Medicine, Zhejiang University. Twenty-two beta- lactamase genes were analyzed by PCR and verified by DNA sequencing. Enzyme activities of extended spectrum beta-lactamases （ESBLs）, cephalosporinase （AmpC） and metallo-beta-lactamases （MBL） were detected by the modified three-dimensional method using enzyme extraction. Results In 62 MDRABs, 51 （82.3%） , 50 （80.6%） and 36 （58.1%） isolates were found to carry blaTEM, blaOXA-23 cluster, and bloADC, respectively. The rest 19 genes were not detected in this study. DNA sequencing and genomic comparison showed that 5 isolates carrying blaTEM had the same genotype as blaTEM-1, 6 isolates carrying blaOXA-23 cluster had the same genotype as blaOXA-23 carbapenemases （accession： CAB69042.1 ） , and 2 isolates carrying blaADC had the same genotype as blaADC-like （accession： EU081908）. Thirty-two isolates （ 51.6% ） produced ESBLs and AmpC, 19 isolates （30.6%） produced ESBLs only, and 1 isolate （ 1.6% ） produced AmpC only; and no isolate produced MBL. Conclusion MDRAB carrying blaOXA-23 carbapenemase and blaADC ArnpC in this study are of high drug resistance.

关 键 词：鲍氏不动杆菌 抗药性 多药 Β-内酰胺酶 基因 酶活性 

分 类 号：R446[医药卫生—诊断学]