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 specific with small amount of sample and short testing time.It is easy to spread in clinical practice.%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。
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