肝癌血清甲胎蛋白免疫磁珠酶联免疫法的建立  

Immunomagnetic Bead Enzyme-linked Immunosorbent Assay Method for Serum AFP in Hepatocellular Cancer Patients

作　　者：邱广亮[1] 赵康[1] 李咏兰[2] 马玉珍 

机构地区：[1]天津大学药物科学与技术学院,天津300072 [2]内蒙古师范大学生物工程研究中心,内蒙古呼和浩特010022 [3]内蒙古医院肿瘤科,内蒙古呼和浩特010017

出　　处：《精细化工》2006年第10期950-953,共4页Fine Chemicals

基　　金：国家自然科学基金资助项目(50066002);内蒙古自然科学基金资助项目(990303-3)~~

摘　　要：采用戊二醛两步交联法将辣根过氧化物酶(HRP)结合于甲胎蛋白(AFP)抗体上制成酶标AFP抗体,质量鉴定结果表明其比活性为200 U/mg,抗体效价为1∶256,满足酶联免疫吸附测定法(ELISA)要求;通过滴定法确定酶标AFP抗体的最适工作稀释度为V(酶标抗体)∶V(磷酸氢二钠-柠檬酸缓冲液)=1∶160;血清用量及孵育时间的影响实验表明:血清100μL、孵育时间2 h为最佳测定条件。建立了肝癌患者血清AFP免疫磁珠ELISA法的规范化检测程序。初步应用实验显示:免疫磁珠ELISA法测定受试健康组血清AFP值平均为3.9 ng/mL,肝癌组血清AFP值平均为316.7 ng/mL,对肝癌诊断阳性率为72.0%。批内及批间CV值分别为5.05%和8.20%。该项技术有较高的灵敏度和很好的特异性,操作简便、分离快速,适用于肝癌的初期快速诊断。Horseradish peroxidase-labelled AFP antibody was prepared using glutaraldehyde two-step crosslinking method. Their specific activity and antibody titer were 200 U/mg and 1：256 respectively. The optimum testing conditions were determined as follows. The working dilution of HRP-labelled AFP antibody was 1 ： 160, the volume of the serum was 100 μL, and the incubation time was 2 h. Immunomagnetic bead enzyme-linked immunosorbent assay （ELISA） method specially for serum AFP in hepatocellular cancer patients was established and used. Preliminary results showed that the positive rate was 72.0% for hepatocellular cancer （HC）. The average value of AFP in the healthy adult group was 3.9 ng/mL,while the average value of AFP in the hepatoeellular cancer group was 316. 7 ng/mL. The CV values within batch and between batches were 5.05% and 8.20% respectively. This technic is not only sensitive and specific but also serves as a simple and rapid means of initial stage diagnosis for hepatocellular carcinoma patients.

关 键 词：肝癌 甲胎蛋白 免疫磁珠 酶联免疫吸附测定法 

分 类 号：Q933[生物学—微生物学]