血清Ⅰ型前胶原放免法的建立和诊断肝纤维化的初步应用

DEVELOPMENT OF RADIOIMMUNOASSAY OF SERUM PROCOLLAGEN TYPE Ⅰ FOR THE DIAGNOSIS OF HEPATIC FIBROSIS AND ITS PRELIMINARY APPLICATION

目的建立测定血清Ⅰ型胶原（ｐｒｏｃｏｌａｇｅｎｔｙｐｅⅠ，ＰＣⅠ）放射免疫分析法（放免法），应用于诊断肝纤维化。方法从流产的人胎皮中提纯Ⅰ型前胶原（ｈｕｍａｎｐｒｏｃｏｌａｇｅｎｔｙｐｅⅠ，ｈＰＣⅠ），免疫兔获特异性抗血清，用改良的氨胺Ｔ法将１２５Ⅰ标记ｈＰＣⅠ抗原，建立双抗体ＰＥＧ法。用此法测定１００例正常人和１４５例各种肝病患者血清ＰＣⅠ。结果建立的放免法标准曲线竞争抑制关系良好，与ＰＣⅢ、ＣⅣ无交叉反应，各种质量指标均达到放免法标准要求。两组肝硬化血清ＰＣⅠ显著升高，而其他各组均无显著升高，血清ＰＣⅠ与ＰＣⅢ水平不相关（ｒ＝０１７２，Ｐ＞００５）。结论用此法测定血清ＰＣⅠ对判断肝纤维化发展至肝硬化有诊断价值。

Objective To develop a radioimmunoassay (RIA) of serum procollagen type Ⅰ(PCⅠ) for the diagnosis of hepatic fibrosis. Methods The antigen human procollagen type Ⅰ(hPCⅠ) was isolated and purified from abored human fetal skin. The specific antiserum was produced from rabbits by immunization used hPCⅠ as antigen. The antigen was labelled with 125Ⅰ by modified chloramine-T method. RIA of double antibody PEG method for determination of serum PCⅠ was developed. Serum PCⅠ of 100 normal controls and 145 patients with varied liver diseases were deterimined by this assay. Results The standard curve had good inhibitory competition of this developed assay, and there was no cross-reactions with procollagen typeⅢ(PCⅢ) and collagen type Ⅳ. All parameters reached the standard of RIA. The levels of serum PCⅠ of two hepatic cirrhotic goups were significantly higher than those of the other groups(P<0 01). There was no cross relationship between serum PCⅠ and PCⅢ (r=0 172, P>0 05). Conclusion The results suggest that the determination of serum PCⅠ used in this assay has significant value in the diagnosis of hepatic fibrosis advancement to cirrhosis.