CA50时间分辨荧光免疫分析法的建立及临床应用

Establishment and clinical application of TRFIA for CA50 measurement

目的建立糖类抗原(CA)50时间分辨荧光免疫分析法(CA50-TRFIA)并用于临床。方法以抗 CA50单克隆抗体包被微孔板,双功能螯合剂异氰酸苄基二乙烯三胺四乙酸(DTTA)络合Eu^(3+)及标记 CA50单克隆抗体,采用夹心法建立 CA50-TRFIA,发光增强系统为以β-二酮体为主的增强液。比较20例消化道恶性肿瘤患者、47例消化道良性病变患者和213名健康对照者的血清 CA50值。数据采用 Logit-Log 函数和四参数 Logistic 函数程序处理。结果方法的批内和批间 CV 分别为2.6%和3.5%,平均回收率为108.61%,灵敏度为1.08×10~3U/L,可测范围为(1.08～140)×10~3U/L。其测定结果与进口的 IRMA 药盒测定值比较,两者高度相关(r=0.904)。对照组213例 CA50测定结果为(2.10±4.96)×10~3 U/L;20例消化道恶性肿瘤 CA50测定结果为(26.58±52.82)×10~3U/L,与对照组比较差异有统计学意义(P<0.01);47例消化道良性病变患者 CA50测定结果为(7.25±28.67)×10~3 U/L,与对照组比较差异无统计学意义(P>0.05)。结论该法简便、快速、实用,精密度、重复性、线性相关性较好。

Objective The aim of this study was to establish a time resolved fluoroimmunoassay （TRFIA） for detecting the carbohydrate antigen （CA） 50 and to evaluate its clinic application. Methods A sandwich TRFIA of CA50 was established, with the monoclonal antibody （McAb） against CA50 coated on the microplate, and the europium（ Eu^3＋ ）-labelled McAb CA50 prepared by the chemical reaction of europium-chelate of N-（ pisothiocyanatobenzyl ）-diethylenetriamine-N, N, N, N-tetraacetic acid. The luminescent enhancement system included a solution mainly containing β-naphthoyl-trifluoroacetone. The serum contents of CA50 in 20 patients with alimentary tract cancers, 47 patients with alimentary carcinoids and 213 normal controls were measured. Results The intra- and inter-assay CV of the TRFIA were 2.6% and 3.5% respectively. The recovery rate was 108.61% , the sensitivity was 1.08 ×10^3 U/L, and the measurement range was （ 1.08 - 140） ×10^3 U/L. There was a high correlation between TRFIA and IRMA （r =0. 904）. The CA50 level was （2.10 24.96） ×10^3 U/L in normal controls, （26.58 250.82） ×10^3 U/L in alimentary tract cancers （ P 〈 0.01, compared with control） and （ 7.25 2 28.67 ） ×10^3 U/L in carcinoids （ P 〉 0. 05, compared with control）. Conclusions The established sandwich TRFIA for CA50 measurement is a simple, fast, practicable, highly specific, sensitive and steady assay. The results of precision, repetition, linear correlation and preliminary clinic application are satisfactory. It is a new non-RIA method for clinic diagnosis and research.