以免疫家兔制备的抗人Ⅳ型胶原多抗包板,采用改良过碘酸钠法辣根过氧化物酶标记抗人Ⅳ胶原单抗,建立了一种血清Ⅳ型胶原的增强化学发光双抗夹心酶免测定法.实验结果表明方法较为灵敏、准确、稳定、特异,其最低检测限为30 μg/L,平均回收率为94.5%,批内和批间变异系数分别≤6.0%和11.6%.测定60例健康人,其参考值为160 μg/L,与进口的日本试剂盒相关性良好,因此可以作为临床血清Ⅳ型胶原放免测定法的参考替代方法.
By utilizing imported chemiluminescent reagent, self-prepared rabbit anti-human type Ⅳ collagen IgG and a horseradish peroxidase-labeled anti-human type Ⅳ collagen monoclonal antibody, a method of sandwich chemiluminescent enzymimmunoassay(CLEIA) was established then evaluated. The results showed the CLEIA has a lower detecting limit of 30 μg/L,the average recovery was 94.5%, CVs of intra-assay and inter-assay were 6.0% and 11.6% respectively, the CLEIA method has a good correlation with Japan kits (ELISA, r=0.9686). the serum reference level by testing 60 cases health is 160 μg/L. It was found that the CLEIA is a sensitive, acurate, stable and specific method,it may be a qualified candidate for serum type Ⅳ collagen detemination of non-radioactive method.
王天成,姚敏捷,刘向祎,寇丽筠.化学发光双抗夹心酶免法测定血清Ⅳ型胶原[J].生物化学与生物物理进展,1998,25(6):554-556
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