人糖链抗原242电化学发光免疫分析检测法的建立

Establishment of electrochemiluminescence immunoassay to detect serum level of CA242

目的建立一种快速、特异、灵敏的检测人糖链抗原242（CA242）的电化学发光免疫分析法（ECLIA）。方法以链霉亲和素包被的磁性微粒、生物素标记的抗CA242单克隆抗体、钌复合物标记配对的抗CA242单克隆抗体组成CA242 ECLIA检测试剂,在ECLIA分析仪上对其特异性、精密度、灵敏度等效能进行方法学评价;用所建立方法与酶联免疫吸附分析法（ELISA）对7例胰腺癌、14例胃癌、25例肠癌患者和20名健康志愿者血清CA242检测结果进行相关性分析。结果所构建的CA242 ECLIA分析法与CA199和CEA无交叉反应,批内精密度为1.4%4.7%,批间精密度为2.5%5.1%,灵敏度为2.0U/mL,测量范围为2.0150.0 U/mL,与ELISA检测结果相关性好（Y=1.066 4＋0.997 9X,r=0.998 9,P〈0.05）,且ELICA检测耗时（18 min）短于ELISA（4 h）。结论本研究所建立的CA242 ECLIA分析法特异性高、灵敏度好、检测速度快、定量检测、无放射性污染,在胰腺癌。

Objective To establish of an electrochemiluminescence immunoassay（ECLIA） to detect serum level of human carbohydrate antigen 242（CA242）.Methods CA242 ELICA kit was composed of biotinylated monoclonal CA242 specific antibody,ruthenium labeled monoclonal CA242 specific antibody and streptavidin-coated microparticles.The performance of established ECLIA,including sensitivity,precision,specificity and so on,was evaluated.The correlation between ECLIA and ELISA in the detection of CA242 was analyzed by using the two methods to detect the serum level of CA242 in 46 cases of patients with gastro-intestinal cancer and 20 healthy contorls.Results No cross-reaction with CA199 and CEA was found.The coefficient of variation of established ECLIA was less than 5.1%,the sensitivity was 2.0 U/mL and the measure range was 2.0150.0 U/mL.There was fine correlation（r=0.998 9,P〈0.05） between ELICA and ELISA.ELICA could be completed in 18 minutes.Conclusion ELICA analysis of CA242,established in this research,could be a specific,sensitive and rapid quantitative method,without radioactive contamination,and could be used for the auxiliary diagnosis and the evaluation of curative effect in patients with cancer