磁分离酶联免疫法检测SCC-Ag的方法学研究  

Methodological study on magnetic affinity immunoassay for detecting SCC-Ag

作　　者：周敏[1] 陆仁飞[2] 杨旻 

机构地区：[1]江苏省南通市疾病预防控制中心,南通226007 [2]南通市第三人民医院 [3]江阴泽成生物技术有限公司

出　　处：《南通大学学报（医学版）》2014年第2期107-110,共4页Journal of Nantong University(Medical sciences)

基　　金：南通市卫生局资助项目(W201224)

摘　　要：目的 ：建立检测鳞状上皮细胞癌抗原（squamous cell carcinoma antigen,SCC-Ag）的磁分离酶联免疫法（magnetic affinity immunoassay,MAIA）。方法：采用双抗体夹心法,用异硫氰酸荧光素（fluorescein isothiocyanate,FITC）和碱性磷酸酶（alkaline phosphatase,AP）分别标记识别不同表位的两种抗SCC-Ag单克隆抗体（mAb）T17和H390,以耦联羊抗FITCmAb包被的免疫磁珠作为固相载体,APS-5作为显色底物,建立检测SCC-Ag的磁分离酶联免疫法。结果：成功实现了用磁分离酶联免疫法对SCC-Ag的定量检测,其检测灵敏度为0.22μg/mL,线性范围0-50μg/mL,批内CV 7%-8%,批间CV 6%-9%。添加回收率为97%。与雅培AxSYM微粒子化学发光检测结果对比,相关系数为0.973。结论：磁分离酶联免疫法的检测结果接近进口同类试剂的检测结果,为开发一种高质量的SSC-Ag测定试剂盒提供实验数据。Objective： To establish magnetic affinity immunoassay（MAIA） for detecting SCC-Ag. Methods： The immunoassay was based on antibody sandwich immunoassay, two monoclonal antibodies T17,H390 were used for conjugating with alkaline phosphatase and with fluorescein isothiocyanate（FITC） respectively, which incorporated magnetic solid phase separation. Magnetic beads were coupled with sheep anti-FITC antibody as solid phase, and APS-5 was used as a substrate to set up MAIA for detecting SCC-Ag. Results： Established the MAIA for detecting SCC-Ag successfully. The sensitivity of the MAIA method was 0.22 μg/mL, the linear range was 0-50 μg/mL, the intra-and inter-assay variation were 7%-8% and 6%-9% respectively. The addition recovery rate was 97%. The correlation coefficient was 0.973 in comparison with Abbott AxSYM. Conclusion： SCC-Ag MAIA kit is better than SCC-Ag RIA and ELISA kit, which can provide experimental data for the development of a high quality of the SSC-Ag assay kit.

关 键 词：鳞状上皮细胞癌抗原 磁分离酶联免疫法 碱性磷酸酶 

分 类 号：R392-33[医药卫生—免疫学]