可溶性细胞间粘附分子-1时间分辨荧光免疫分析法的建立及对肺癌诊断的临床应用

Establishment of TRFIA for soluble intercellular adhesion molecule-1 and Its Clinical Application in Diagnosis of Lung Cancer

目的建立可溶性细胞间粘附分子-1生物素-亲和素系统(BSA)时间分辨荧光免疫分析方法 ,并对其方法学及对肺癌诊断临床应用价值进行评价。方法用2株匹配的单克隆抗体分别作为捕获抗体和检测抗体,利用制备的铕标记链亲和素(SA-Eu3+)作为示踪物并与生物素化的检测抗体特异结合,建立双位点多层夹心法BSA-TRFIA法,检测68例正常对照者和61例肺癌患者血清sICAM-1水平,对其方法学和临床应用价值进行评价。结果 sICAM-1TRFIA检测灵敏度为1.32μg/L,批内和批间CV分别为4.24%和6.83%,平均回收率为99.87%,与ELISA法测定值的相关系数为0.939,可测范围为2.5-933μg/L,肺癌组与健康对照组比较具有显著性差异(P<0.01)。结论 sICAM-1时间分辨荧光免疫分析(TRFIA)是一种新型非放射性标记免疫分析法,具有良好的特异性、准确度、稳定性,可作为肺癌的辅助诊断指标。

Objective Soluble intercellular adhesion molecule-1（sICAM-1） is an predictive marker for inflammation events.The purpose of our study was to establish a time-resolved fluoroimmunoassay（TRFIA） for sICAM-1-biotin-streptavidin complex and evaluate its diagnostic and clinical application in lung cancer.Methods A two-sided multi ＂sandwich＂ immunoassay biotin-streptacidin（BSA）,time-resolved fluoroimmunoassay（TRFIA） was developed.In conjunction with pre-synthesized SA-Eu3＋ specifically bound to biotin detection antibody,it was used as a marker substance for detection of sICAM-1 level in plasma.The serum levels of sICAM-1 in 68 patients with lung cancer and 61 normal controls were assayed by TRFIA and the results were compared with those by ELISA using correlation analysis.Results The lower detection limit was 1.32 μg/L.The intra-and inter-assay CVs were 4.24% and 6.83%,respectively.The mean recovery rate was 99.87%.The linear range of the methods for sICAM-1detection was from 2.5 to 933 μg/L.The results measured by the TRFIA and ELISA methods were closely correlated（R2=0.939）.There was significantly difference between the group of patients with lung cancer and the one of normal controls in the serum levels of sICAM-1（P0.01）.Conclusion The TRFIA for sICAM-1 measurement is an exceedingly specific,accurate and steady assay.It is radiation-free also,and it can be served as an auxiliary index in the diagnosis of lung cancer.