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.
出处
《现代检验医学杂志》
CAS
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
