【摘要】 目的建立磁微粒化学发光免疫分析法检测血清Ⅰ型胶原蛋白C末端交联肽(CTX)。方法将异硫氰酸荧光素(FITC)标记的抗体R1、碱性磷酸酶标记的配对抗体R2(R1、R2为抗CTX单克隆抗体)和待测标本结合形成复合物。复合物通过抗FITC抗体特异性结合到磁微粒上,在磁场作用下,复合物与未结合的其他物质分离。加入酶促化学发光底物,底物发生酶促反应,形成发光反应,在检测范围内,发光强度与标本中的CTX水平成正比。评价所建立方法的最低检测限、精密度、线性范围、回收率等性能指标。结果所建立的检测方法线性范围为0.01～6.00ng/mL,在此线性范围内,r≥0.990;检测限为0.009 8ng/mL;批内与批间精密度分别为2.6%～5.9%和4.1%～8.1%;回收率在98.9%～102.6%;检测结果与罗氏公司生产的CTX试剂盒的检测结果具有显著相关性。结论该研究所建立的磁微粒化学发光免疫分析法用于检测血清CTX能够满足临床需求,具有较好的推广运用价值。更多还原

【Abstract】 Objective To establish magnetic particle chemiluminescence immunoassay for detecting serum collagen typeⅠ C-terminal cross-linked peptide(CTX).Methods Fluorescein isothiocyanate(FITC)labeled antibody R1 and alkaline phosphatase labeled antibody R2(R1 and R2 were anti-CTX monoclonal antibodies)were combined with the detected specimen to form a complex.The complex was specifically bound to magnetic particle by anti-FITC antibody,under the action of magnetic field,the complex was separated from other substances which were not bound.When enzyme catalyzed chemiluminescence substrate was added,the substrate reacts enzymously to form the luminescent reaction,within the detection range,the luminous intensity was directly proportional to the CTX level in the specimen.The minimum detection limit,precision,linear range and recovery rate of the method were evaluated.Results The linear range of the method was 0.01-6.00 ng/mL,and r≥0.990 in this linear range.The detection limit was 0.009 8 ng/mL.The intra assay and inter assay precisions were 2.6%-5.9%and 4.1%-8.1%respectively.The recovery rates were in the range of 98.9%-102.6%.There was a significant correlation between the detection results and the results of CTX kit produced by Roche.Conclusion The magnetic particle chemiluminescence immunoassay established by the research could meet clinical needs for detecting serum CTX,and has a good value for promotion and application.更多还原