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投稿时间：2019-11-17 网络发布日期：2020-09-20
投稿时间：2019-11-17 网络发布日期：2020-09-20
中文摘要: 目的 通过检测分析耐碳青霉烯肺炎克雷伯菌(CRKP)的流行病学资料,研究耐药趋势及耐药机制。方法 收集山西白求恩医院2018年1月至2019年1月临床分离的耐碳青霉烯肺炎克雷伯菌,对其进行鉴定、药敏及表型筛选。通过PCR方法检测主要碳青霉烯耐药基因［A类丝氨酸酶 (KPC),B类金属β-内酰胺酶(NDM型、IMP型、VIM型)以及D类丝氨酸酶(OXA)-48等］和外膜孔道蛋白基因(OmpK35、OmpK36),并对孔道蛋白基因检测的阳性菌株采用实时荧光定量PCR进行基因表达的相对定量分析。结果 经分析，CRKP的分离率为4.91%。CRKP时间分布以2019年1月分离率最高,为29.27%;CRKP年龄主要集中在21~80岁,其中以41~50岁分离率最高,为24.39%;科室分布以重症医学科为主,分离率为24.39%,其次为普通外科21.95%、康复医学科14.63%及神经外科12.20%;标本分布以痰标本为主,分离率为31.71%,其次为尿19.51%、血19.51%及分泌物12.20%。CRKP对亚胺培南、厄他培南的耐药率分别为90.24%、100.00%;对环丙沙星及阿米卡星的耐药率分别为95.12%和56.10%,对β-内酰胺类、氨基糖苷类耐药率均较高。改良碳青霉烯灭活实验(mCIM)和EDTA-改良碳青霉烯灭活实验(eCIM)显示有31株(75.61%)菌株表型表现为丝氨酸酶,有3株(7.32%)为金属酶;检出的碳青霉烯酶基因,有39株(95.12%)为KPC型,1株(2.44%)NDM 型,1株(2.44%)同时具有以上KPC型和NDM型;只有1株(2.44%)菌株缺失OmpK36基因,余均存在OmpK35及OmpK36。进一步分析OmpK35中41株只有9株(21.95%)表达下调,而OmpK36中40株有38株(95.00%)表达下调。结论 CRKP耐药现象严峻,感染年龄以中青年为主,且主要集中在重症医学科,以痰液标本为主;其CRKP的耐药机制主要为产KPC型丝氨酸酶和外膜孔道蛋白OmpK36缺失及表达下降。
Abstract:Objective To investigate the trend and mechanism of drug resistance of carbapenem-resistant Klebsiella pneumoniae(CRKP) by the epidemiological data. Methods The CRKP isolated from Shanxi Bethune Hospital from January 2018 to January 2019 were collected for identification, drug sensitivity test and phenotype screening.The major carbapenem-resistance genes ［class A (KPC), class B (NDM, IMP, VIM), class D (OXA-48)］ and outer membrane pore protein (OmpK 35, OmpK 36) were detected by PCR.The relative quantitative analysis of gene expression was performed by real-time quantitative PCR. Results The separation rate of CRKP was 4.91%.The highest separation rate of CRKP was 29.27% in January 2019.The age distribution of CRKP was mainly concentrated in 21-80 years old, with the highest separation rate of 24.39% between 41-50 years old.The distribution of departments was mainly in the ICU with the separation rate of 24.39%, followed by General Surgery 21.95%, Rehabilitation Medicine 14.63% and Neurosurgery 12.20%;the distribution of specimens was mainly sputum samples, the separation rate was 31.71%, followed by urine 19.51%, blood 19.51% and secretion 12.20%.The drug resistance rates of CRKP to imipenem and ertapenem were 90.24% and 100.00%, respectively;the drug resistance rates to ciprofloxacin and amikacin were 95.12% and 56.10%, respectively.The resistance rates to β-lactamases and aminoglycosides were higher.The results of modified carbapenem inactivation method (mCIM) and EDTA modified carbapenem inactivation method (ECIM) showed that 31 strains (75.61%) were serinase phenotype, 3 strains (7.32%) were metalloenzyme;39 strains (95.12%) were KPC type, 1 strain (2.44%) was NDM type, and 1 strain (2.44%) had both KPC type and NDM type. Only one strain (2.44%) had OmpK36 gene deletion, and the others had OmpK35 and OmpK36.There were 9 (21.95%) of 41 of OmpK35 were down-regulated, while 38 (95.00%) of 40 of OmpK36 were down regulated. Conclusion The drug resistance of CRKP is severe.The infection age was mainly young and middle-aged, and mainly concentrated in the ICU.The drug resistance mechanism of CRKP was mainly the production of KPC type serinase, and the loss and decreased expression of OmpK36.
keywords: Klebsiella pneumoniae Carbapenem antibiotics Resistance trend Resistance mechanism Klebsiela pneumoniae carbapenemase Outer membrane pore protein
文章编号： 中图分类号： 文献标志码：A
基金项目:山西省应用基础研究项目(201601D011116)
| Author Name | Affiliation |
| SONG Jing,RONG Jian-rong,HOU Chen-rui | Laboratory Department,Shanxi Bethune Hospital,Taiyuan,Shanxi 030032,China |
引用文本：
宋静,戎建荣,侯辰蕊.耐碳青霉烯肺炎克雷伯菌41株分布及耐药机制[J].中国临床研究,2020,33(9):1163-1166,1171.
宋静,戎建荣,侯辰蕊.耐碳青霉烯肺炎克雷伯菌41株分布及耐药机制[J].中国临床研究,2020,33(9):1163-1166,1171.