夹心酶联免疫吸附试验检测人表皮生长因子的方法建立及其应用研究

目的:建立临床适用的表皮生长因子(epidermalgrowthfactor,EGF)酶联免疫检测方法,探讨EGF在临床常见肿瘤患者血清中的含量变化规律。方法:采用EGF单抗包被酶标板,兔抗EGF多抗为二抗,以辣根过氧化物酶标记的羊抗兔IgG作为酶标抗体建立夹心法ELISA检测临床常见肿瘤患者及健康体检人群的血清EGF含量。结果:建立的EGF酶联免疫吸附法最小检测限达15.6ng/L,批内精密度为8.7%,批间精密度为11.6%,平均回收率为94.7%,符合临床检测要求。血清EGF含量在31名正常人为(1.305±0.382)ng/mL,24名乳腺癌病人术前值为(0.729±0.347)ng/mL,术后为(0.711±0.332)ng/mL,17名结直肠癌病人术前为(0.678±0.311)ng/mL,术后为(0.658±0.298)ng/mL,19名胃癌病人术前为(0.598±0.224)ng/mL,术后为(0.614±0.257)ng/mL。各肿瘤组患者血清EGF水平较健康对照组明显降低(P<0.01)。结论:建立了适用于临床检测的EGF酶联免疫检测方法,肿瘤患者血清中EGF含量较健康对照组明显下降,该检...

Establishment of measuring serum EGF by enzyme-linked immunosorbent assay and its clinical application

Objective To develop enzyme-linked immunosorbent assay(ELISA)for quantitation of endogenous human epidermal growth factor (EGF) and to detect immunoreactive concentration of EGF in patients with carcinoma. Methods The present assay systems were based on the sandwiching of the antigen between a monoclonal mouse anti-hEGF antibody,pre-coated on a 96-well polystyrene plate,and a polyclone rabbit anti-hEGF antibody,which is then detected with a peroxidase-labeled goat anti-rabbit antibody. Results The calibrati...