癌胚抗原的时间分辨免疫荧光分析及其诊断试剂的研制

Time-resolved fluoroimmunoassay of carcino-embryonic antigen and preparation of it’s diagnostic reagent

目的：利用时间分辨免疫荧光分析（TRFIA）技术，建立一种人血清癌胚抗原（CEA）的快速全自动检测方法并研制其诊断试剂。方法：采用双抗体夹心法（用1株抗CEA的mAbM86160M用于包被微孔板，另1株抗CEA的mAb M86110M用于标记铕）建立CEA—TRFIA，增强液采用B-二酮体为主的发光增强系统。结果：CEA-TRFIA的线性测量范围为（1～560）μg／L，分析的灵敏度为0．28μg／L，批内和批间的变异系数（CV）分别为7．2％～8．6％和8．9％～13．2％。与AFP，CA12—5、CA19—9和白蛋白均无交叉反应，与CA15—3的交义反应值为1．62μg／L。将1000份血清标本用本法与国外罗氏化学发光试剂盒同时检测，其相关系数（r）为0．946。结论：CEA—TRFIA的各项指标均达到临床检测的要求，可替代国外同类检测试剂盒，用于临床血清CEA水平的测定。

AIM： TO establish a two-site time-resolved fluoroimmunoassay （TRFIA） for detection of carcino-embryonic antigen （CEA） based on the direct sandwich technique （One monoclonal antibody（mAb） is immobilized on the surface of microtiter plate strip wells, another mAb is labelled with Eu^3＋ ） and prepare its diagnostic reagent. METHODS： The sandwich TRFIA for detection of serum CEA level using anti-CEA mAbs. RESULTS： The measure range of CEATRFIA was （ 1 - 560） μg/L. The analytical sensitivity was 0. 28 μg/L. The intraand inter-assay coefficients of variation（CV） were 7.2% -8.6% and 8.9% - 13.2%, respectively. There was no cross-reaction to AFP, CA12-5, CA19- 9 and albumin . Cross reactivity to CA15-3 was 1. 62 μg/L. The correlation coefficient of blood samples detected by CEA-TRFIA and commercially available CEA CLIA（ chemiluminescent immunoassay） kit （Roche） was 0. 946. CONCLUSION： CEA-TRFIA is a new immunoassay which has several advantages including high sensitivity, specificity and a wide working range. It is suitable for routine clinical use.