肝纤维化早期定量诊断方法研究

目的：建立一种敏感、特异、操作简单的血清Ⅳ型胶原蛋白（ＣＯＬⅣ）测定方法，并可与肝炎标记物同步进行测定。方法：以免疫家兔制备抗人ＣＯＬⅣ多抗包被微孔板，采用改良过碘酸钠法辣根过氧化物酶标记抗人ＣＯＬⅣ单抗，改变各种稀释液成分及浓度，建立双抗夹心酶免疫测定法（ＥＬＩＳＡ）。结果：最低检测极限为 ３０μｇ／Ｌ，平均回收率为１０２．７５％，批内和批间变异系数分别为＜６．０％和１１．８％；测定５０例健康人，其参考值为９７．９μｇ／Ｌ，测定２７例肝纤维化病人血清平均值为２５４．８２μｇ／Ｌ。结论：此方法具有灵敏、准确、稳定、特异性较高、标本用量少、检验用时短，并可与乙型肝炎标志物同时检测的特点，易在临床推广使用。

The research on measurement Type

Objective To establish a sensitive, specific and simple measurement for serum collagen Ⅳ which can be used with HBsAg, HBsAb. HBeAb HBcAg and HbcAb. Method By Preparing anti - human type Ⅳ collagen IgG with rabbits and utilizing sodium periodate horse - radish peroxidase - labelling anti - human type Ⅳ collagen monoclonal antibody to change dilution component and concentration, we established sandwich ELISA. Results ELISA had a lowest testing limit of 30μg/L, the average recovery was 102. 75%, CVs of intra - assay were respectively 6.0 and 11.8% and the reference value by testing 50 healthy people was 97.9μg/L±36.3μg/L, the average value of liver cirrhosis type Ⅳ collagen was 254.82μg/L±69.9μ/g. Conclusion The improved method is sensitive, exact, stable and spe- cific with small amount of sample and short testing time. It is easy to spread in clinical practice.