CA242高灵敏时间分辨荧光免疫分析及其初步应用  

A high sensitivity time resolved fluoroimmunoassay of CA242 and its preliminary clinical application

作　　者：黄飚[1] 盛世乐[2] 朱岚[1] 刘海燕[1] 陆云[3] 张珏[1] 谢敏浩[1] 

机构地区：[1]江苏省原子医学研究所,无锡214063 [2]上海交通大学医学院附属仁济医院核医学科 [3]复旦大学附属华山医院核医学科

出　　处：《中华核医学杂志》2009年第4期268-270,共3页Chinese Journal of Nuclear Medicine

基　　金：江苏省自然科学基金（BK020133）;江苏省卫生厅科研项目（H200856）

摘　　要：目的采用时间分辨荧光免疫分析技术（TRFIA）建立高灵敏的恶性肿瘤相关糖类抗原（CA）242的快速全自动检测方法并进行初步应用。方法以抗CA242单克隆抗体601#包被96孔微孔板，以Eu^3＋-N^2-[p-异氰酸苄基]-二乙烯三胺四乙酸标记抗CA242单克隆抗体242#，发光增强系统为以β-二酮体为主的增强液。采用平衡饱和法建立CA242 TRFIA，采用Auto DELFIA1235系统考核CA242 TRFIA，并进行健康人和胃癌患者血清样本的检测。结果该方法的批内和批间CV分别为2．92％和5．36％，平均回收率为98．75％，灵敏度为0．2kU／L，可测范围为0．2～200kU／L，最高浓度点计数的20％、50％、80％对应的效应点值（ED20、ED50、ED80）分别为25．2kU／L、53．3kU／L和112．5kU／L。CA50、CA19-9对CA242分别有15％和32％的交叉反应，癌胚抗原（CEA）、甲胎蛋白（AFP）、CA125对CA242 TRFIA均无交叉反应。Eu^3＋标记242#抗体于-20℃下保存6个月免疫反应性基本无损，同批试剂连续6个月应用分析结果稳定，检测健康体格检查人员的血清，样品的平均值为（6．52±5．15）kU／L，建议阳性阈值为17kU／L，临床结果与酶联免疫吸附测定（ELISA）的结果相符，r为0．916。116例胃癌患者血清CA242的阳性率为26．7％。结论CA242 TRFIA的灵敏度高、稳定性好，可用于血清CA242含量的免疫测定。Objective CA242 is valuable in the screening, diagnosis and prognosis prediction in pancreatic and colorectal tumors. The aim of this study was to establish a two-site time-resolved fluoroimmunoassay （TRFIA） of CA242 based on the sandwich technique and to study its clinical usefulness. Methods By the equilibrium method, the monoclonal antibodies （McAb） directed against a specific antigenic site on the CA242 was No. 601#, and the europium labelled McAb prepared from the europium chelate N-（ p-isothiocyanatobenzyl）-diethylenetriamine-N,N, N, N-tetraacetic acid directed against a different antigenic site on the CA242 molecule was No. 242#. The luminescent enhancement system contained mainly 2-naphthoyltrifluoroacetone. CA242 in sera from patients and healthy controls were determined by CA242-TRFIA by auto DELFIA1235 system. Results The intra- and inter-assay CV of the CA242-TRFIA were 2.92% and 5.36% , respectively, and the recovery rate was 98.75%. The sensitivity was 0.2 kU/L and the measurement range was 0.2-200 kU/L. The cross-reacting rates with CA50 and CA19-9 were 15% and 32% , respectively, while those with carcino-embryonic antigen （ CEA）, alpha fetoprotein （AFP） and CA125 were negligible. The europium labelled No. 242# was at least stable for six months at - 20℃. Serum samples from healthy blood donors were analyzed, resulting in a mean value of （6.52 ±5.15） kU/L, with an estimated reference limit ≤17 kU/L. The linear correlation of TRFIA and ELISA measurements resulted in a correlation coefficient of 0. 916. The observed positive rate of CA242 in 116 gastric cancer cases was 26. 7%. Conclusion The newly developed CA242-TRFIA technique was highly sensitive and stable in the immuno-determination of serum CA242.

关 键 词：荧光免疫测定 抗原 肿瘤相关 碳水化合物 消化系统肿瘤 

分 类 号：R686[医药卫生—骨科学]