糖类抗原125化学发光免疫定量检测试剂的研制与临床应用

Development and Clinical Application of a Chemiluminescence Immunoassay for Determination of Carbohydrate Antigen 125

摘要: 为了研制肿瘤标志物糖类抗原125(CA125)化学发光免疫定量检测试剂，并分析其临床应用价值，将CA125单抗包被微孔板，以辣根过氧化物酶标记的CA125抗体为二抗，结合鲁米诺化学发光底物系统，建立CA125化学发光免疫定量检测方法。用CA125检测试剂国家参考品质控血清分析所建立方法的特异性、灵敏度、稳定性和精密性等试剂性能，并与进口的CA125定量检测试剂同时检测临床血清样本350例，比较检测结果。结果表明所研制的试剂性能符合CA125检测试剂国家参考品质量标准。批内变异系数2.8%~3.6%、批间变异系数3.7%；试剂盒置37℃孵育3d，其稳定性良好。临床样本比对检测结果表明，两种方法相关性良好(相关系数r = 0.9897，阴性符合率99.5%、阳性符合率99.3%，总符合率为99.4%，Kappa值为0.99，一致性强度为最强)。本文成功研制了快速、特异、敏感和稳定的肿瘤标记物CA125化学发光免疫定量检测试剂并建立其检测方法，适合临床检验推广应用。

Abstract: To develop a chemiluminescence immunoassay (CLIA) kit for quantitative determination of tumor marker carbohydrate antigen 125 (CA125) and evaluate its value of clinical application, CLIA for quantitative determination of CA125 was established by using micro-plates coated with antibody to CA125, combined with HRP conjugated anti-CA125 antibody, as well as luminol chemiluminescence substrate system. The national reference control sera were used for evaluation of specificity, sensitivity, stability and accuracy of the established CLIA. CA125 levels in sera from 350 clinical samples were parallel determined by the established methods and reference kits produced by the Siemens. The results show that the efficiency of the CLIA reagents met the requirements of national reference standard. The intra coefficient variations (CV) were between 2.8%~3.6% and inter CV 3.7%. The kit showed good stability after being kept at 37˚C for 3d. The established CLIA showed good correlation with the reference kits (negative concordance rate, positive concordance rate and overall concordance rate were 99.5%, 99.3% and 99.4% respectively, and a correlation coefficient r = 0.9897. High degree of agreement was observed with a Kappa value of 0.99). A rapid, specific, sensitive and stable CLIA kit for quantitative determina-tion of CA125 was successfully established and that will be fit for widely application in clinical laboratory.