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目的：建立血清Ⅳ型胶原蛋白（COLW）浓度测定的酶联免疫试验（ELISA）方法，辅助诊断肝硬化。方法：制备过氧化物酶（HRP）-免抗人COLⅣ多抗结合物，作为标记抗体；用棋盘滴定试验确定包被抗体和标记抗体浓度，进行检出限、精密度、回收率、特异性、灵敏度测定和标准曲线的制备；初步用于79例正常人和34例肝纤维化患者。结果：COLⅣ浓度在40~I000μg／L之间时，该法工作曲线最佳，最低检出限为40μg／L，批内异变系数（CV）6．098％，批间CV4．30％，高、低浓度回收率分别为95％和98％。该法测定正常人COLⅣ浓度平均为67.6±24.6μg／L，而肝纤维化患者平均水平为215.96％±96μg／L，可见辅助诊断肝纤维化具有很高特异性和灵敏度。结论：ELISA方法检测定血清COLⅣ简便、特异、准确、价格低廉，可用于肝纤维化辅助诊断并在国内推广应用。
Objective: To establish an enzyme - linked immunosorbent assay (ELISA) for the determination of serum concentration of type Ⅳ collagen (COLW) to assist in the diagnosis of liver cirrhosis. METHODS: Peroxidase (HRP) -anti-anti-human COLIV polyclonal antibody was prepared and used as a labeled antibody. The concentration of coated antibody and labeled antibody was determined by checkerboard titration. The detection limit, precision, recovery, specificity , The determination of sensitivity and the preparation of standard curve; initially used in 79 normal subjects and 34 patients with liver fibrosis. Results: When the concentration of COL Ⅳ was between 40 ~ 1000μg / L, the best curve was obtained. The detection limit was 40μg / L, the coefficient of variation (CV) 6.098%, the inter-assay CV 4.30% High and low concentration recovery were 95% and 98%. The method for determination of normal COL Ⅳ concentration average 67.6 ± 24.6μg / L, while the average level of liver fibrosis 215.96% ± 96μg / L, can be seen auxiliary diagnosis of liver fibrosis with high specificity and sensitivity. CONCLUSION: ELISA assay is a simple, specific, accurate and inexpensive method for the determination of COL Ⅳ. It can be used in the diagnosis of hepatic fibrosis and its application in China.