纹带棒状杆菌耐药特征研究  

Research on drug resistance characteristics of Corynebacterium striatum

作　　者：魏孔娇 王嘉正 邱小彤 徐帅[3] 朱雄[4] 李欢[4] 王晓霞 王成玲 范仕弘 韩李超 刘志国[3] 李振军 WEI Kongjiao;WANG Jiazheng;QIU Xiaotong;XU Shuai;ZHU Xiong;LI Huan;WANG Xiaoxia;WANG Chengling;FAN Shihong;HAN Lichao;LIU Zhiguo;LI Zhenjun(Department of Medicine,Tibet University,Lasa 850000,Tibet,China;Department of Clinical Laboratory Medicine,The First Affiliated Hospital of Shandong First Medical University&Shandong Provincial Qianfoshan Hospital,Shandong Medicine and Health Key Laboratory of Laboratory Medicine,Jinan 250000,Shandong Province,China;National Institute for Communicable Disease Control and Prevention,Chinese Center for Disease Control and Prevention,Beijing 102206,China;Central and Clinical Laboratory of Sanya People’s Hospital,Sanya 572000,Hainan Province,China)

机构地区：[1]西藏大学医学院,西藏自治区拉萨850000 [2]山东第一医科大学第一附属医院(山东省千佛山医院)检验医学科,山东省医药卫生临床检验诊断学重点实验室,山东济南250000 [3]中国疾病预防控制中心传染病预防控制所,传染病预防控制国家重点实验室,北京102206 [4]海南省三亚市人民医院检验科,中心实验室,海南三亚572000

出　　处：《微生物与感染》2021年第6期384-390,共7页Journal of Microbes and Infections

基　　金：国家自然科学基金(82073624);国家重点研发计划(2019YFC1200601,2019YFC1200705);海南省自然科学基金青年基金(818QN326)。

摘　　要：分析我国山东地区临床分离的纹带棒状杆菌对常用药物的敏感性,并对其耐药机制进行探讨。收集该省某三甲医院临床样本,用哥伦比亚血平板培养基对纹带棒状杆菌进行分离培养,用微量肉汤稀释法检测12种常用抗生素对纹带棒状杆菌的最低抑菌浓度(minimal inhibitory concentration,MIC)。采用聚合酶链式反应(polymerase chain reaction,PCR)对耐药菌株的耐药基因[aph(3”)-Ⅰb、aph(6)-Ⅰd、aac(6’)-Ⅰb,tetW,ermX,gyrA]进行扩增、测序比对,以探讨相关的耐药机制。通过实时荧光定量PCR(quantitative real-time PCR,qPCR)对耐药基因ermX、tetW进行定量分析,采用公式2^(-ΔΔCt)计算基因相对表达量差异倍数。结果显示,共分离出纹带棒状杆菌83株,且均为多重耐药菌。对万古霉素、利奈唑胺表现为完全敏感;对红霉素、环丙沙星、克林霉素表现出完全耐药,对四环素的耐药率为86.7%(72/83),对庆大霉素的耐药率为38.6%(32/83);ermX基因的检出率为100%(83/83),tetW基因的检出率为88.0%(73/83),aph(3”)-Ⅰb基因的检出率为36.1%(30/83),aph(6)-Ⅰd基因的检出率为37.3%(31/83),aac(6’)-Ⅰb基因的检出率为15.7%(13/83)。ermX和tetW基因的相对表达量均有所变化,但变化不明显。内在型耐药(cMLS)是菌株对红霉素耐药的潜在机制。结果提示,本研究调查医院的纹带棒状杆菌耐药严重,耐药谱较为广泛,须引起医院和实验室的重视。To reveal the drug resistance and its mechanism preliminarily of clinical isolates of Corynebacterium striatum in Shandong Province of China,83 isolates of Corynebacterium striatum were collected from clinical specimens during March 2017 and June 2020 in a tertiary hospitals from Shandong.The minimal broth dilution method was used to detect the minimal inhibitory concentration.The aminoglycoside modifying enzyme genes[aph(3”)-Ⅰb、aph(6)-Ⅰd、aac(6’)-Ⅰb],the ribosomal protective protein gene of tetracycline resistance(tetW),the ribosomal methylase gene(ermX),quinolone resistance genes(gyrA)were tested by polymerase chain reaction(PCR).The PCR products were sequenced and compared with BLAST.Quantitative real-time PCR(qPCR)was used to analyze the drug-resistant genes ermX and tetW quantitatively,and formula 2-ΔΔCt was used to calculate the relative expression of genes.All the tested Corynebacterium striatum were proved as multi-drug resistant strains.The MDR-Cs showed completely sensitive to vancomycin and linezolid and completely resistance to erythromycin,ciprofloxacin and clindamycin,the detection rate of tetracycline,gentamicin were 86.7%(72/83)and 38.6%(32/83),respectively.The detection rate of ermX and tetW were 100%(83/83)and 88%(73/83),the detection rate of aph(3”)-Ⅰb,aph(6)-Ⅰd,aac(6’)-Ⅰb were 36.1%(30/83),37.3%(31/83)and 15.7%(13/83),respectively.The resistance phenotype of the strain to erythromycin is cMLS.The drug resistance spectrum is relatively broad in this study,which needs to be paid attention from hospitals and laboratories.

关 键 词：纹带棒状杆菌 药物敏感性 耐药基因 实时荧光定量PCR 

分 类 号：R378[医药卫生—病原生物学]