Arakawa H, Ikegami T, Maeda M, Tsuji A
School of Pharmaceutical Sciences, Showa University, Tokyo, Japan.
J Biolumin Chemilumin. 1993 May-Jun;8(3):135-9. doi: 10.1002/bio.1170080302.
We developed a sensitive chemiluminescent sandwich-type enzyme immunoassay (CL-EIA) of alpha-fetoprotein (AFP) using beta-D-galactosidase (beta-gal) as a label and 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside as a substrate. The CL-EIA for AFP was performed using two monoclonal antibodies, one antibody is labelled with beta-gal, the other is coated onto the inside surface of a polystyrene tube. The detection limit for AFP was 0.5 ng/mL, equivalent to 10 pg/assay tube. The coefficient of variation for within and between assay imprecision were 2.0%-4.9% (n = 10) and 4.4%-9.8% (n = 5), respectively. AFP values in serum determined by this method correlated well with those obtained by radioimmunoassay (n = 26, r = 0.99). This sensitive AFP assay can be performed within 4 h and can be used as a routine assay in clinical diagnosis.
我们开发了一种灵敏的化学发光夹心型酶免疫分析法（CL-EIA）用于检测甲胎蛋白（AFP），该方法使用β-D-半乳糖苷酶（β-gal）作为标记物，5-溴-4-氯-3-吲哚基-β-D-吡喃半乳糖苷作为底物。AFP的CL-EIA使用两种单克隆抗体进行，一种抗体用β-gal标记，另一种抗体包被在聚苯乙烯管的内表面。AFP的检测限为0.5 ng/mL，相当于每检测管10 pg。批内和批间精密度的变异系数分别为2.0%-4.9%（n = 10）和4.4%-9.8%（n = 5）。用该方法测定的血清中AFP值与放射免疫分析法得到的值相关性良好（n = 26，r = 0.99）。这种灵敏的AFP检测法可在4小时内完成，可作为临床诊断的常规检测方法。
