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基于上转磷光颗粒的AFP免疫层析定量检测
引用本文:赵露晶,周蕾,方菲,孙红英,杨瑞馥,沈鹤柏.基于上转磷光颗粒的AFP免疫层析定量检测[J].生物磁学,2009(15):2907-2909.
作者姓名:赵露晶  周蕾  方菲  孙红英  杨瑞馥  沈鹤柏
作者单位:[1]上海师范大学生命与环境科学学院,上海200234 [2]军事医学科学院微生物流行病研究所,北京100071
基金项目:基金项目:上海市科学技术委员会(0752nm028,0952nm05500和08TC140600)项目资助
摘    要:目的:建立了上转磷光免疫层析快速定量检测AFP的方法,用于原发性肝癌的诊断。方法:利用上转磷光标记的双抗夹心免疫层析技术检测血清中AFP的含量。将AFP单克隆抗体分别标记上转磷光颗粒并包被硝酸纤维膜,制成免疫层析检测试纸。评价该试纸的灵敏度、特异性和精密度。并通过临床50例血清样本,探讨上转磷光技术与化学发光检测方法的一致性。结果:该方法能在20min内完成,检测灵敏度达5ng/ml。浓度范围从5-1000ng/ml作标准曲线,呈现较好的线性。通过三组平行实验探讨其精密度,变异系数都在10%以内。应用于其它肿瘤标志物如CEA、CA199、AFU和NSE的检测,无交叉反应。与化学发光相比,决定系数R2=0.9692,一致性较好。结论:该方法简便、快速、廉价,可以实现定量,尤其适合基层门诊和体检现场使用。

关 键 词:上转磷光技术  免疫层析测定  甲胎蛋白  定量测定

Development of up-converting phosphor immunochromatography test for quantitative detection of AFP
Institution:ZHAO Lu-jing, ZHOU Lei, FANG Fei, SUN Hong-ying, YANG Rui-fu, SHEN He-hai (1 College of Environmental Science, Shanghai Norma University, Shanghai 200234, China; 2 Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China)
Abstract:Objective: To develop a method for rapid and quantitative detection of AFP based up-converting phosphor technology. Method: An Up-Converting Phosphor immunochromatography test based on double- ntibody sandwich assay has been developed for de- tection of AFP. The anti-AFP monoclonal antibody is conjugated to the up-converting phosphor particles while the same antibody immo- bilized on the nitrocellulose membrancc. We evaluated the sensitivity, specificity and precision of this method. The concordance rate be- tween UPT and Axsym was discussed through 50 clinical sera. Results: Typical analysis time was less than 20 minutes. And the sensitivity of the test was 5 ng/ml. When the standard quantitative curve was determined, a good linearity was displayed in the range from 5 to 1000 ng/ml. The precision of the assay was below 10%(coefficient of variation, CV), and no cross reaction was found when the method was used to detect other tumor marker (including CEA, CA199, AFU and NSE). Compared to the method of Axsym by determination coeffi- cient (R2=0.9692), it shows good correlation. Conclusion: The method is simple, rapid, cheap and quantitative, especially suitable for e- mergency and screening on physical examination.
Keywords:Up-converting phosphor technology  Immunchromatographic assay  AFP  Quantitative detection
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