碳青霉烯类耐药革兰阴性菌对黏菌素和多黏菌素B的药敏试验方法学评价  

Evaluation of methods for testing the susceptibility of carbapenem-resistant gram-negative bacilli to colistin and polymyxin B

作　　者：郭燕 尹丹丹 胡付品 高晶晶 何丽华[3] 吴文娟[3] 孙景勇[4] 倪语星[4] 汤瑾 余佳佳[6] 刘瑛[6] 田东兴 张泓 刘文健[8] 赵虎[8] 李敏[2] GUO Yan;YIN Dandan;HU Fupin;GAO Jingjing;HE Lihua;WU Wenjuan;SUN Jingyong;NI Yuxing;TANG Jin;YU Jiajia;LIU Ying;TIAN Dongxing;ZHANG Hong;LIU Wenjian;ZHAO Hu;LI Min(Institute of Antibiotics,Huashan Hospital,Fudan University,Shanghai 200040,China)

机构地区：[1]复旦大学附属华山医院抗生素研究所,上海200040 [2]上海交通大学医学院附属仁济医院 [3]上海同济大学附属东方医院南院 [4]上海交通大学医学院附属瑞金医院 [5]上海交通大学附属第六人民医院 [6]上海交通大学医学院附属新华医院 [7]上海交通大学附属儿童医院 [8]复旦大学附属华东医院

出　　处：《中国感染与化疗杂志》2020年第5期525-535,共11页Chinese Journal of Infection and Chemotherapy

基　　金：上海市科学技术委员会“科技创新行动计划”产学研医合作项目(17DZ1910403);上海市细菌真菌耐药监测网(3030231003)。

摘　　要：目的评估各种药敏试验方法检测碳青霉烯类耐药菌株对黏菌素和多黏菌素B敏感性结果的可靠性。方法以肉汤微量稀释法为参比方法,采用E试验法、VITEK 2-Compact法、纸片扩散法、MIC测试条(MTS)和梳状条(E strip)法5种药敏试验方法测定1040株碳青霉烯类耐药的肺炎克雷伯菌(CRKP)、铜绿假单胞菌(CRPA)和鲍曼不动杆菌(CRAB)对黏菌素和多黏菌素B的敏感性。结果E试验法、VITEK 2-Compact法和纸片扩散法测定377株CRKP对黏菌素的敏感率分别为97.1%、96.3%和98.1%;317株CRPA的敏感率分别为95.3%、100%和99.1%;346株CRAB的敏感率分别为100%、99.7%和99.7%。MTS、E strip法和纸片扩散法测定CRKP对多黏菌素B的敏感率分别为97.3%、97.3%和99.2%;317株CRPA的敏感率分别为98.7%、99.4%和99.7%;346株CRAB的敏感率分别为100%、100%和99.7%。与肉汤微量稀释法测定结果相比,5种方法检测黏菌素或多黏菌素B的分类一致率(CA)均>95%。对于CRKP,5种方法的非常重大误差(VME)为1/9~5/7,重大误差(ME)为0.3%~66.7%;对于CRPA菌株,E试验法和MTS法的基本一致率(EA)<90%,5种方法均未检出VME;E试验法检测黏菌素的ME为0.3%~17.6%;对于CRAB菌株,除VITEK 2-Compact法外其他4种方法的VME为1/3~1/1,ME为0~0.3%。结论E试验法、VITEK 2-Compact法、纸片扩散法、MTS和E strip法等检测黏菌素和多黏菌素B对碳青霉烯类耐药革兰阴性菌的药敏,均存在EA偏低或假敏感现象(即VME),需引起实验室和临床高度重视,实验室仍应按美国临床和实验室标准化协会(CLSI)文件采用标准肉汤微量稀释法进行多黏菌素药敏试验。Objective To evaluate the accuracy of five methods for testing the in vitro susceptibility of carbapenem-resistant gram-negative bacteria to colistin and polymyxins B.Methods Antimicrobial susceptibility was determined for a total of 1040 isolates of carbapenem-resistant Klebsiella pneumoniae(CRKP),Pseudomonas aeruginosa(CRPA)and Acinetobacter baumannii(CRAB)using Etest,VITEK 2-Compact,disk diffusion,MIC Test Strips(MTS)and E strip.Broth microdilution was taken as reference method.Results Etest,VITEK 2-Compact,and disk diffusion method resulted in 97.1%,96.3%and 98.1%of the 377 CRKP strains,95.3%,100%and 99.1%of the 317 CRPA strains,100%,99.7%and 99.7%of the 346 CRAB strains susceptible to colistin,respectively.MTS,E strip,and disk diffusion method reported 97.3%,97.3%and 99.2%of the CRKP strains,98.7%,99.4%and 99.7%of the CRPA strains,100%,100%and 99.7%of the CRAB strains susceptible to polymyxin,respectively.Taking broth microdilution method as reference,the five comparative testing methods reported categorical agreement(CA)higher than 95%in the susceptibility results of colistin and polymyxin B.The five methods also reported very major error(VME)from 1/9 to 5/7,and major error(ME)from 0.3%to 66.7%,in the results for CRKP.Etest and MTS reported lower essential agreement(EA)and no VME in the results for CRPA strains.Etest method reported ME from 0.3%to 17.6%in the results of colistin against CRPA strains.In the susceptibility results of CRAB strains,the other four methods except VITEK 2-Compact showed VME from 1/3 to 1/1 and ME from 0 to 0.3%.Conclusions Etest,VITEK 2-Compact,disk diffusion,MTS,and E strip have issues such as low rate of essential agreement or false sensitivity(VME)in testing the susceptibility of carbapenem-resistant K.pneumoniae,A.baumannii and P.aeruginosa to colistin and polymyxin B,which is a concern for laboratory staff and clinicians.Laboratories should still adopt standard broth microdilution method for testing the susceptibility of K.pneumoniae,A.baumannii and P.aeruginosa to colistin a

关 键 词：黏菌素 多黏菌素B 碳青霉烯类 药敏试验方法学 

分 类 号：R978[医药卫生—药品]