两种碳青霉烯酶检测方法的临床应用研究  

Clinical application research of two carbapenemase detecting methods

作　　者：傅俊方 谭伟清[1] 林子沛 丁新 刘婉婷 江凌晓[1] FU Junfang;TAN Weiqing;LIN Zipei;DING Xin;LIU Wanting;JIANG Lingxiao(Division of Laboratory Medicine,Zhujiang Hospital of Southern Medical University,Guangzhou,Guangdong 510282,China;Department of Laboratory Medicine,the People's Hospital of Gaoming District of Foshan,Foshan,Guangdong 528500,China)

机构地区：[1]南方医科大学珠江医院检验医学部,广东广州510282 [2]佛山市高明区人民医院检验科,广东佛山528500

出　　处：《热带医学杂志》2023年第12期1654-1658,1695,共6页Journal of Tropical Medicine

基　　金：国家重点研发计划(2017YFC1200801)

摘　　要：目的评估碳青霉烯酶抑制剂增强试验和碳青霉烯耐药基因检测试验分析肠杆菌目细菌产碳青霉烯酶性能和临床应用价值。方法收集2021年2月-2022年6月南方医科大学珠江医院从临床样本中分离的耐碳青霉烯肠杆菌目细菌(CRE)共372株(非重复分离株),分别用碳青霉烯酶抑制剂增强试验分析其碳青霉烯酶酶型,用荧光PCR法检测5种碳青霉烯耐药基因(bla_(KPC)、bla_(NDM)、bla_(IMP)、bla_(VIM)、bla_(OXA-48)),比较两种检测方法的结果。结果372株CRE分离株主要分离自重症医学科病区143株(其中综合ICU 73株,急诊科ICU 38株),占分离总数的38.4%;神经内科病区47株,占分离总数的12.6%。碳青霉烯酶抑制剂增强试验检测结果显示,产A类丝氨酸碳青霉烯酶250株,产B类金属β内酰胺酶96株,有26株为不产A类丝氨酸碳青霉烯酶或B类金属β内酰胺酶。荧光PCR法共检测出携带碳青霉烯耐药基因的有333株(89.5%),其中携带bla_(KPC)基因的有234株(62.9%),携带bla_(NDM)基因的有96株(25.8%),携带bla_(IMP)、bla_(KPC+NDM)、bla_(OXA-48)基因的各有1株,未检出携带bla_(VIM)基因的菌株。两种检测方法相比较,酶抑制剂增强试验和荧光PCR法对单产A类丝氨酸碳青霉烯酶的检测结果分别为250、234株,对单独产B类金属β内酰胺酶的检测结果分别为96、97株;两种检测方法在2种酶型检测结果的一致性检验Kappa值分别为0.788、0.797,具有良好的一致性(P均<0.05)。结论酶抑制剂增强试验和荧光PCR法具有良好的一致性、操作简便,结果易于判读、互为补充,具有较高的临床检测效能。Objective To evaluate the efficiency and application value of carbapenemase inhibitor-enhancing assays and carbapenems-resistant genes detection test in the detection of carbapenemase in carbapenem-resistant Enterobacterales(CRE).Methods Totally 372 strains(non-repetitive isolated colony)of CRE isolated from clinical specimens of patients at the Zhujiang Hospital of Southern Medical University from February 2021 to June 2022 were collected.The carbapenemase was detected by carbapenemase inhibitor-enhancing assays and fluorescent polymerase chain reaction was used to detect five kinds of carbapenems-resistant genes including bla_(KPC),bla_(NDM),bla_(IMP),bla_(VIM)and blaOXA-48.The results of the two detection methods were analyzed and compared.Results The 372 CRE isolates were mainly isolated from the ICU ward with 143 isolates,accounting for 38.4%of the total isolates(including comprehensive ICU 73 strains and 38 strains in the emergency department)and the neurology ward with 47 isolates,accounting for 12.6%of the total isolates.The result of carbapenemase inhibitor enhancing assays showed that 250 strains produced Class A serine carbapenemase,96 strains produced Class B metallo-β-lactamase,and 26 strains produced neither Class A serine carbapenemase nor Class B metallo-β-lactamase.A total of 333 strains carrying carbapenemase genes were detected by fluorescent polymerase chain reaction,accounting for 89.5%.Among them,234 strains carried the bla_(KPC)gene,accounting for 62.9%.There were 96 strains carrying the bla_(NDM)gene,accounting for 25.8%of the total,and one strain each carried the bla_(IMP)gene,bla_(KPC+NDM)genes,and blaOXA-48 gene.No strains carried the bla_(VIM)gene were detected.Comparing these two detecting methods,the detection results for the production of Class A serine carbapenemases using carbapenemase inhibitor enhancing assays and fluorescent polymerase chain reaction were 250 and 234 strains,respectively.The detection results for the production of Class B metallo-β-lactamase using these two

关 键 词：耐碳青霉烯肠杆菌目细菌 碳青霉烯酶抑制剂增强试验 荧光PCR法 碳青霉烯酶检测 

分 类 号：R378[医药卫生—病原生物学]