荧光微球时间分辨免疫层析技术定量检测甲胎蛋白的研究  

Quantitation of alpha fetoprotein(AFP) in human serum using fluorescence microspheres time-resolved immunochromatographic technology

作　　者：郭明明[1] 周衍[1] 周剑波[2] 俞静文[2] 陈燕[2] 范俊[1] 黄飚[1] 

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]江阴市人民医院检验科,214400

出　　处：《免疫学杂志》2015年第10期897-901,共5页Immunological Journal

基　　金：江苏省卫生计生委科研课题(Z201506)

摘　　要：目的建立荧光微球时间分辨免疫层析技术检测血清中甲胎蛋白（AFP）的反应体系并对该法进行评价。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的单克隆抗体AC18#,将抗AFP单克隆抗体AC17#和羊抗鼠抗体喷涂于硝酸纤维素膜上分别作为检测线和质控线,制作免疫层析试纸条。结果标准曲线经Log-Log B函数处理,检测AFP的线性范围为0.03~1 000 U/ml,批内和批间变异系数（CV）分别为5.35%和8.94%,平均回收率为98.45%。该法检测100例健康人员的AFP水平为（2.04±1.92）U/ml,正常参考值为5.80 U/ml,与时间分辨免疫分析法（TRFIA）检测AFP有较好的相关性,r=0.978（P〈0.05）。结论本研究建立的荧光微球时间分辨免疫层析具有快速、灵敏、简便、经济、可单人份操作等优点,为检测AFP提供了一种高效的新途径,符合我国国情,便于临床推广。This study was designed to develop a fluorescent microspheres time-resolved immunochromatographicassay for detection of alpha fetoprotein（AFP） in human serum, and to evaluate the method performance. The Eu-time-resolved fluorescent polystyrene particle conjugated with the monoclonal antibody AC18#was used as a fluorescent label. The monoclonal antibody AC17#and goat anti-mouse Ig G were immobilized on the nitrocellulose membrane as the test line and control line. The linear response range of the method to detect the serum AFP was 0.03-1 000 U/ml. And the intra- and inter- assay coefficient were 5.35% and 8.94%,respectively, while average recovery rate was 98.45%. The means of 100 healthy volunteers was（2.04 ±1.92） U/ml,thus the normal ranges of serum AFP levels were less than 5.80 U/ml. Measurements obtained by the fluorescent microspheres time-resolved immunochromatographic strip showed a correlation coefficient（r） of 0.978（P 0.05） with measurements obtained by TRFIA. In conclusion, the fluorescent microspheres time-resolved immunochromatographic strip for assaying AFP provided high sensitivity, rapid, simple operation, low cost, single-servings and can be extensively used for clinical operations.

关 键 词：荧光微球 时间分辨 免疫层析 甲胎蛋白 

分 类 号：R446.6[医药卫生—诊断学]