人血清中总前列腺特异性抗原化学发光免疫分析法的建立

Establishment of Chemiluminescent Immunoassay for Measurements of Total-Prostate Specific Antigen in Human Serum

目的建立一种定量分析人血清中总前列腺特异性抗原（t—PSA）的化学发光免疫分析方法。方法采用生物素和碱性磷酸酶分别标记捕获抗体与检测抗体，与样本中的抗原形成免疫复合物，然后通过链霉亲和素与生物素的反应被链霉亲和素标记的磁微粒捕获，利用碱性磷酸酶与发光底物的反应建立了双单抗夹心法检测人血清中t—PSA的化学发光免疫分析方法。结果标准品对WHO96／670国际标准品进行溯源，同时检测血清样本，线性回归方程为y=0．9434X-0．1605，相关系数r=0．99。选择具有等摩尔分子反应性的单抗，对不同存在形式的PSA检测偏差在士3％以内。病人血清样本的测试结果与雅培architecti2000化学发光试剂盒检测结果显著相关，相关系数r=0．989。结论该方法具有较高的灵敏度和重复性，检测结果准确，提高了国产同类试剂的竞争力，同时比同类进口试剂成本低，适于临床检测和科研应用。

Objective To establish chemiluminescent immunoassay for the quantitative measurement of total prostate specific antigen （t-PSA） in the human serum. Methods The immunoassay utilized biotin which could react with streptavidin linked with magbeads to label with capture antibody and used alkaline phosphotase as the labeled enzyme,to be with the antigen in sample,it could form the immuno-complex. Through the reaction between biotin and streptavidin, the complex was fixed on the magbeads. The substrate would react with alkaline phosphotase and the double sandwich chemiluminescent immunoassay was established. Results Trace to WHO 96/670 reference materials,in-house standard materials had been established,the results of serum correlated well with WHO 96/670,with linearity equation Y= 0. 943 4X-0. 160 5, correlation coefficient was 0.99. The paired monoclonal antibody with equimolar characteristics was selected,the detection deviation with different forms of PSA was in the range of ±3%. The serum tested results correlated well with that of AB- BOTT architect i2 000 CLIA kit and the correlation coefficient was 0. 989. Conclusion With the consideration of highly sensitivity,repeatition and auuracy,the competence of the reagents would be promoted. Simuhaneously,the reagents are low-cost and are suitable for clinical application and scientific research.