表皮生长因子酶联免疫吸附检测方法的建立  

Establishment of the Standard Curve of Measuring Serum EGF by Indirect Competitive Enzymelinked Immunosorbent Assay

作　　者：吴学诗[1] 夏勇[1] 李雪玲[2] 张美英[2] 

机构地区：[1]广州医学院附属第二人民医院检验科,广东广州510150 [2]广州暨南大学生物医药研发基地,广东广州510632

出　　处：《现代检验医学杂志》2004年第4期4-6,共3页Journal of Modern Laboratory Medicine

基　　金：广东省重大攻关课题立项 粤科计字 [2 0 0 0 ] 2 61

摘　　要：目的　建立酶联免疫吸附法 ( ELISA)检测人血清 (浆 )或尿液中表皮生长因子的含量。方法　以rh EGF为包被抗原 ,表皮生长因子 ( epidermal growth factor,EGF)为竞争的抗原 ,两者与一定量的抗EGF多抗反应。以抑制率为纵坐标 ,EGF的对数值为横坐标建立标准曲线。结果　实验结果表明 ,理想的包被抗原浓度为 1μg/ml,抗 EGF多抗工作浓度为 1∶ 1 0 0 0 0 0 ,酶标二抗工作浓度为 1∶ 30 0 0 ,可测量最适范围为 0 .2 5～ 32 ng/ml,最小检测量为 0 .5 ng/ml,批内和批间变异系数分别为 3.48%和 5 .46%。得到回归方程 Y=- 0 .1 2 81 Ln( X) + 0 .81 1 6( r2 =0 .9938)和标准曲线。结论　建立快速定量测定Objective To establish the ELISA to detect the concentration of EGF in serum or urine samples.Methods EGF was man-made coating antigen.The EGF was an competitor to rhEGF.They could react to the limited amout of polyclonal antibody against EGF.Results The result of experiment showed that the optimal concentration of the coating antigen,polyclonal antibody against EGF and sheep anti-rabbit IgG were 1 μg/ml,1∶100 000 and 1∶3 000 respectively.Quantization of the EGF was linear from 0.25～32 ng/ml,with the detection limit being 0.5 ng/ml.The coefficients of variation of intra-assay and inter-assay were 3.48% and 5.46% respectively.Then,the regression equation was Y=-0.128 1 Ln(X)+0.8116(r 2=0.993 8) and there was a standard curve.Conclusion Indirect competitive enzyme-linked immunosorbant assay is established to measure rapidly the serum EGF.The whole assay can be completed within 6 hours.

关 键 词：EGF CIELISA 标准曲线 

分 类 号：R446.61[医药卫生—诊断学]