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目的对临床分离的碳青霉烯类耐药肺炎克雷伯菌(CR-KPN)进行多位点序列分型(MLST),同时研究不同ST分型感染患者的临床特点。方法收集2013年1月-2015年6月辽宁省为主5所医院临床分离的61株CR-KPN,采用微量肉汤稀释法测定受试菌株对14种抗菌药物的敏感性。应用PCR基因扩增技术及DNA测序方法对受试菌株进行碳青霉烯酶基因检测。通过对受试菌株进行MLST,探讨其克隆相关性和分子流行病学特征。同时分析不同ST分型下CR-KPN的耐药特点、耐药机制及菌株感染的临床特点。结果 MLST分型显示61株CR-KPN共有18种ST分型,ST11为优势型别(53.3%),研究同时发现这部分ST11型菌株更容易对碳青霉烯类抗生素耐药且MIC值较高,大部分产KPC-2型碳青霉烯酶。单因素分析提示ST11组患者在ICU接受治疗所占的比例及机械通气的使用率高于非ST11组,差异具有统计学意义。其他ST型中ST2033、ST2135、ST2193、ST2194、ST2195、ST2196为世界范围内首次注册,其中ST2193、ST2194、ST2195与ST11有密切的亲缘性。临床资料提示同一所医院同一时期以相同克隆流行为主。结论 MLST显示CR-KPN共有18种ST型别,ST11为优势型别,结合临床资料发现同一所医院同一时期以相同克隆流行为主,提示CR-KPN有局部播散的风险。鉴于临床资料分析提示接受过ICU治疗和机械通气的患者容易发生CR-KPN菌株(特别是ST11型)感染的风险,此类患者应引起临床的广泛重视。
Objective To investigate the clinical features of clinically isolated carbapenem-resistant Klebsiella pneumoniae (CR-KPN) by multilocus sequence typing (MLST). Methods Sixty-one CR-KPN strains isolated from five hospitals in Liaoning Province from January 2013 to June 2015 were collected. The susceptibility of the tested strains to 14 antimicrobial agents was determined by the broth microdilution method. The carbapenemase gene was tested on the tested strains by PCR amplification and DNA sequencing. The MLST of the tested strains was used to investigate the clonal correlation and molecular epidemiological characteristics. At the same time, we analyzed the drug resistance, drug resistance mechanism and clinical characteristics of CR-KPN in different ST types. Results There were 18 ST-types in 61 CR-KPNs and 53.3% ST-STs in 61 strains of CR-KPN. The results showed that ST11-type strains were more susceptible to carbapenem antibiotics and MIC values High, mostly produced KPC-2 carbapenemase. Univariate analysis showed that the proportion of ST11 patients receiving ICU and the rate of mechanical ventilation were higher than those of non-ST11 group, the difference was statistically significant. ST2033, ST2135, ST2193, ST2194, ST2195 and ST2196 are the first registered in the world in other ST types, of which ST2193, ST2194 and ST2195 are closely related to ST11. Clinical data suggest that the same hospital during the same cloning-based prevalence. Conclusions There are 18 ST types in CR-KPN and ST11 is the predominant type in MLST. According to the clinical data, the same clinic prevalence in the same hospital in the same period suggests that CR-KPN has the risk of local spread. Given that clinical data analysis suggests that patients with ICU-treated and mechanically ventilated patients are at high risk for developing CR-KPN strains (particularly ST11), such patients should attract widespread clinical attention.