糖类抗原CA19-9光激化学发光免疫分析方法的建立

目的:建立一种检测人血清糖类抗原CA19-9(CA19-9)浓度的光激化学发光免疫分析方法。方法:采用2株针对不同抗原表位的抗CA19-9抗体,其中一株抗体用来包被发光微粒,另一株抗体标记生物素,以双抗体夹心法检测血清中CA19-9浓度,并与Roche公司试剂(电化学发光法)进行比较。结果:发光微粒浓度为25 mg/ml、生物素标记抗体浓度为16.17 mg/ml时,监测范围为0 U/ml～500 U/ml,灵敏度为0.27 U/ml,平均回收率为101.26%,批内变异系数(CV)为1.67%～4.35%,批间CV为1.12%～3.25%,与CEA、CA12-5、CA15-3的交叉反应率低,稳定性好,与电化学发光免疫分析法的符合率高(符合率=98.8%,γ=0.991)。结论:光激化学发光免疫分析方法测定CA19-9具有高灵敏度、精密度和准确性,与电化学发光免疫分析法的符合率高,适用于临床测定。

Development of light initiated chemiluminescent immunoassay for Carbohydrate antigen CA19-9

Objective:To develop a light initiated chemiluminescent immunoassay for detection of Carbohydrate antigen CA19-9(CA19-9) in human serum.Methods: Two monoclonal antibodies with different epitopes to CA19-9 were selected.One was coated with luminescent microspheres,and the other was biotinylated.The CA19-9 concentration in human sera was determined by double antibody sandwich light initiated chemiluminescent immunoassay,and the results were compared with Roche Reagent(electro-chemiluminescent immunoassay).Results: The detection range of the assay was 0 U/ml~500 U/ml,when the concentrations of the luminescent microspheres and the biotinylated antibody were 25 mg/ml and 16.17 mg/ml respectively.The sensitivity was 0.27 U/ml,and the recovery rate was 101.26%.The within-run and between-run coefficients of variation(CV)were 1.67%~4.35% and 1.12%~3.25% respectively.The cross reaction rates of the assay with carcino-embryonic antigen(CEA),carbohydrate antigen 12-5(CA12-5),carbohydrate antigen 15-3(CA15-3) were low.The stability of the detection method was good.The coincidence rate with the electro-chemiluminescent immunoassay was highly significant(conformity=98.8%,γ=0.991).Conclusion: The light initiated chemiluminescent immunoassay for human CA19-9 detection have high sensitivity,precision and accuracy.The coincidence rate with Roche kits is high,so this assay can be applied for clinical determination.