胶体金免疫层析法应用于快速检测CRE的碳青霉烯酶  

Application of colloidal gold immunochromatography in rapid determination of carbapene⁃mase in CRE

作　　者：罗国兰 李梦薇 张飞龙 朱虹贞 林盛张 陈丽坤 蒋利君 王海霞 鲁炳怀 韦柳华 LUO Guolan;LI Mengwei;ZHANG Feilong;ZHU Hongzhen;LIN Shengzhang;CHEN Likun;JIANG Lijun;WANG Haixia;LU Binghuai;WEI Liuhua(Department of medical laboratory,Liuzhou WorkersHospital,Liuzhou,Guangxi,China,545005;Peking University China-Japan Friendship School of Clinical Medicine,Beijing,China,100029;Laboratory of Clinical Microbiology and Infectious Diseases,Department of Respiratory and Critical Care Medicine,China-Japan Friendship Hospital,Beijing,China,100029)

机构地区：[1]柳州市工人医院医学检验科,广西柳州545005 [2]北京大学中日友好临床医学院,北京100029 [3]中日友好医院呼吸与危重症医学科临床微生物与感染实验室,北京100029

出　　处：《分子诊断与治疗杂志》2024年第12期2342-2346,共5页Journal of Molecular Diagnostics and Therapy

基　　金：广西壮族自治区卫生健康委员会自筹经费科研课题(Z20210030)。

摘　　要：目的评估胶体金免疫层析法(GICA)快速检测耐碳青霉烯类肠杆菌目细菌(CRE)的碳青霉烯酶的临床应用价值。方法对2016年3月至2021年11月柳州地区非重复的CRE临床分离株54株,采用碳青霉烯酶表型实验初步筛查,多重实时定量PCR复核。上述方法无法明确耐药基因的菌株,采用WGS进行检测,同时GICA快速检测CRE的碳青霉烯酶类型。以多重实时定量PCR为参考方法,评估GICA快速检测CRE的碳青霉烯酶的准确性。结果本次收集的CRE菌株为54株,主要是大肠埃希菌(27株,50.0%)、肺炎克雷伯菌(14株,25.9%)。碳青霉烯酶表型实验结果显示单产金属酶的CRE有50株(92.6%),单产丝氨酸酶的CRE有2株(3.7%),同时产金属酶与丝氨酸酶的CRE有2株(3.7%)。GICA结果与多重实时定量PCR的检测结果一致,显示54株CRE中有50株(92.6%)NDM阳性,2株(3.7%)KPC阳性,1株(1.9%)NDM+KPC阳性,1株(1.9%)NDM+OXA⁃48阳性。GICA在快速检测CRE碳青霉烯酶类型与多重荧光定量PCR的结果一致性为100%。GICA对NDM、KPC及OXA⁃48阳性菌株酶型检测的准确性均为100%。结论GICA在检测CRE碳青霉烯酶中具有快速简便及准确度高的优点,助力临床精准治疗CRE感染具有重要的应用价值。Objective To evaluate clinical application value of colloidal gold immunochromatogra⁃phy(GICA)for rapid detection of carbapenemase in carbapenem⁃resistant Enterobacteriaceae(CRE)isolates.Methods Totally,54 non⁃reduplicative isolates of CRE were collected from March 2016 to November 2021 in L iuzhou.The expression of carbapenemase gene was preliminary screening by the carbapenemase phenotype test and the review determination of carbapenemase by multiple real⁃time quantitative PCR.For the strains whose drug resistance genes could not be identified by the above methods,the whole genome sequencing(WGS)was used for detection.The type of carbapenemase in CRE was rapidly detected by GICA.Multiple real⁃time quantita⁃tive PCR was used as reference method to evaluate the accuracy of GICA in the rapid determination of carbapen⁃emase.Results The CRE collected in this study were 54 isolates,mainly including Escherichia coli(27 isolates,50.0%)and Klebsiella pneumoniae(14 isolates,25.9%).The results of carbapenemase phenotype test showed that there were 50 metal enzymes isolates(92.6%),2 serine enzymes isolates(3.7%),and 2 both metal enzymes and serine enzymes isolates(3.7%).Consistent with the detection results of multiple real⁃time quantita⁃tive PCR,the results of colloidal gold immunochromatography,showed that there were 50 NDM positive isolates(92.6%),2 KPC positive isolates(3.7%),1 NDM+KPC positive isolate(1.9%)and 1 NDM+OXA⁃48 positive isolate(1.9%).The consistency of colloidal gold immunochromatography and multiplex fluorescent quantitative PCR in rapid detection of carbapenemase types of CRE isolates was 100%.The accuracy of GICA for the deter⁃mination of the carbapenemase types of NDM,KPC and OXA⁃48 positive isolates was 100%.Conclusion GICA has the advantages of rapid,simple and high accuracy in rapid determination of carbapenemase in CRE,which has important application value in clinical precise treatment of CRE infection.

关 键 词：耐碳青霉烯类肠杆菌目细菌 碳青霉烯酶 快速检测 胶体金免疫层析法 多重实时定量PCR 

分 类 号：R446.6[医药卫生—诊断学]