时间分辨荧光免疫层析法联合检测血清肿瘤标志物  

Simultaneous detection of tumor markers in human serum using time-resolved fluorescence microspheres immunochromatographic assay

作　　者：郭明明[1] 肖华龙[2] 周剑波[3] 傅利新 周彬[1] 张珏[1] 黄飚[1] Guo Mingming;Xiao Hualong;Zhou Jianbo;Fu Lixin;Zhou Bin;Zhang Jue;Huang Biao(Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi 214063, China;Department of Clinical Laboratory, Wuxi People＇s Hospital, Wuxi 214023, China;Department of Clinical Laboratory, Jiangyin People＇s Hospital, Jiangyin 214400, Chin;Wuxi Jiangynan Industrial Technology and Trade Corporation. Wuxi 214063. China)

机构地区：[1]江苏省原子医学研究所、卫生部核医学重点实验室、江苏省分子核医学重点实验室,无锡214063 [2]无锡市人民医院检验科,214023 [3]江阴市人民医院检验科,214400 [4]无锡市江原实业技贸总公司,214063

出　　处：《中华核医学与分子影像杂志》2018年第4期266-270,共5页Chinese Journal of Nuclear Medicine and Molecular Imaging

基　　金：江苏省科技支撑计划-社会发展（BE2014608）

摘　　要：目的建立时间分辨荧光免疫层析法（TRFMIA）联合检测血清甲胎蛋白（AFP）和癌胚抗原（CEA）的反应体系，并对其进行评价。方法以包裹有Eu螯合物的荧光微球作为标记物，共价偶联抗AFP单克隆抗体（简称单抗）AC18^#和CEA单抗AE03^#，将抗AFP单抗AC17^#、抗CEA单抗AE05^#和羊抗兔IgG抗体喷涂于硝酸纤维素膜上，作为检测线和质控线，制作免疫层析试纸条，确定检测方法线性范围、最低检测限等指标，并将该法与时间分辨荧光免疫法（TRFIA）和电化学发光免疫分析法（ECLIA）进行直线相关分析。结果标准曲线显示，AFP可测范围为0.07～1 000.00 kU/L，批内和批间CV分别为5.93%和11.07%，平均回收率为92.77%；CEA可测范围为0.12～500.00 μg/L，批内和批间CV分别为7.53%和12.13%，平均回收率为94.73%。TRFMIA与TRFIA和ECLIA检测的AFP和CEA结果有较好的相关性，相关系数为0.93～0.97。结论成功建立时间分辨荧光免疫层析试纸联合检测法，可同时检测AFP和CEA。ObjectiveTo develop a time-resolved fluorescent microspheres immunochromatographic assay （TRFMIA） for detection of alphafetoprotein （AFP） and carcinoembryonic antigen （CEA） in human serum and to evaluate its performance.MethodsThe Eu-time-resolved fluorescent polystyrene particles conjugated with monoclonal antibody AC18^# for AFP and AE03^# for CEA were used as fluorescent labels. The monoclonal antibody AC17^# for AFP, AE05^# for CEA and goat anti-rabbit antibody were immobilized on the nitrocellulose membrane as the test lines and control line. Several performances indicators were measured, including linear range, detection limit, and specificity. AFP and CEA were measured by the new method and the results were compared with those obtained by time-resolved fluoroimmunoassay （TRFIA） and electro-chemiluminescence immunoassay （ECLIA） using linear correlation analysis.ResultsThe measurement ranges of AFP were 0.07-1 000.00 kU/L with the intra- and inter-assay CV of 5.93% and 11.07%, and those of CEA were 0.12-500.00 μg/L with the intra- and inter-assay CV of 7.53% and 12.13% respectively. The average recovery rate of AFP and CEA was 92.77% and 94.73%, respectively. Measurements obtained by TRFMIA had strong correlation coefficients ranging from 0.93 to 0.97 when compared with those obtained by TRFIA and ECLIA.ConclusionTRFMIA, which can simultaneously detect AFP and CEA, has been successfully established.

关 键 词：甲胎蛋白类 癌胚抗原 荧光免疫测定 化学发光测定法 

分 类 号：R730.43[医药卫生—肿瘤]