电化学酶免疫分析法测定甲胎蛋白  

Electrochemical Enzyme Immunoassay for the Detection of Human Alpha-fetoprotein (AFP)

作　　者：郑莉[1] 胡荫华[1] 宋俊峰[1] 邓贤碧[2] 梁国栋[2] 

机构地区：[1]西北大学化学系,西安太白路1号710069 [2]第四军医大学附属二院中心实验室,西安市长乐路710032

出　　处：《西北大学学报（自然科学版）》1993年第1期23-29,共7页Journal of Northwest University（Natural Science Edition）

基　　金：国家自然科学基金

摘　　要：将酶联免疫吸附试验(ELISA)与单扫描示波极谱(LSP)相连接,使用直接与酶联板配套的小型三电极系统,建立了测定甲胎蛋白(AFP)的新方法——电化学酶免疫分析法。测得AFP浓度与极谱峰峰高在0.5～100ng/mL范围内呈良好线性关系(r=0.9984),检出限为0.0625ng/mL,比ELISA测得值灵敏约一个数量级。测定了36名健康人血清中AFP含量,结果与文献报道相符。测定了32例病人血清标本中AFP含量,并将结果与ELISA试验测得值进行比较,经统计学处理,两者无显著性差异(t=1.291,P>0.05)。AFP, a fetal protein, was used in the diagnosis and follow—up to liver cancer and in prenatal diagnosis of fetal malformations as well as for monitoring the effect of therapy. In this paper, a sandwich enzymelinked immunosorbent assay utilizing linear sweep polarography detection (ELISA LSP) for AFP in human serum was established successfully. A smaller three electrode systerm (DME Pt Ag/AgCl) was designed to insert directly in the well on commercial ELISA plate, which simplified the procedure, enhanced the sensitivity and lowered the detection error by ELISA—LSP. Horaseradish peroxidase (HRP) as labelling enzyme catalyzes conversion of o phenylenediamine to 2. 2' diaminoazobenzene(DAA). At pH9～11, well defined cathodic peak of DAA at Ep 800 mV(vs. Ag/AgCl) was achieved. Under optimum conditions, the linear relation between polarographic (second derivative) peak height of DAA and AFP concentration was ranged 0.5 to 100ng/mL with r=0.9984. The detection limit of AFP was 0.0625ng/mL, which was 10 times more sensitive than that by ELISA. The AFP levels of 36 specimens of healthy human serum detected are 1.2 to 12.8 ng/mL with the average value of 4.6±2.2 ng/mL, which agreed with the data reported in literature. The clinical evaluation of AFP levels of 32 patient specimens by ELISA—LSP and by current ELISA using spectrophotometric detection was compared. The results obtained were in good agreement (t=1.291, P>0.05).

关 键 词：抗体 极谱分析 电化学 甲胎蛋白 

分 类 号：Q51[生物学—生物化学]