可溶性细胞间黏附分子-1时间分辨荧光免疫分析法的建立  

Time-resolved fluoroimmunoassay for soluble intercellular adhesion molecule-l-biotin-streptavidin complex

作　　者：肖华龙[1] 黄飚[2] 张晓峰[1] 虞竞峰[1] 陆国础[1] 朱岚[2] 强新晨[1] 祝炳方[1] 郁震[1] 

机构地区：[1]江苏省无锡市人民医院检验科,214023 [2]江苏省无锡市人民医院卫生部核医学重点实验室,214023

出　　处：《中华核医学杂志》2008年第5期344-346,共3页Chinese Journal of Nuclear Medicine

基　　金：基金项目：卫生部核医学重点实验室开放课题基金（WK005-021）

摘　　要：目的建立一种新的高灵敏度、宽范围的可溶性细胞间黏附分子-1（sICAM-1）生物素．链亲和素（BSA）系统时间分辨荧光免疫分析（TRFIA）方法并用于临床。方法使用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—Eu^3＋）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并进行质量控制，考核其灵敏度、特异性、测量范围及方法稳定性等。检测32例血清样品中sICAM-1含量，并与ELISA所测结果进行相关分析。测定31名健康对照者和22例肺部疾病患者血样中的sICAM-1含量。结果该法测定sICAM-1的灵敏度为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％，线性范围为（2．5～1933）μg/L。该方法测定值与ELISA所测结果高度相关（R^2=0．939）。与肿瘤坏死因子（TNF）-α、白细胞介素（IL）-8、IL-1、IL-2和IL-6等无交叉反应。溶血、脂血和高胆红素等血清标本对该方法基本无干扰。31名健康对照者测定结果为（365．74±45．03）μg／L。22例肺部疾病患者血样结果示，在炎症和肺损伤的情况下，可见sICAM-1有升高的趋势。结论该sICAM-1 BSA-TRFIA法灵敏度高，可测范围广，可以方便地应用于临床。Objective The soluble intercellular adhesion molecule-1 （sICAM-1） is an effective marker for inflammation. The purpose of this study was to establish a novel, highly sensitive, wide range and time-resolved fluoroimmunoassay for sICAM-1-biotin-streptavidin complex for use in clinical practice. Methods Two coupled monoclonal antibodies were used as capture and detection antibodies. A two-sided multi ＂sandwich＂ immunoassay biotin-streptavidin （ BSA）, time-resolved fluoroimmunoassay （TRFIA） was developed. In conjunction with pre-synthesized SA-Eu^3＋ specifically bound to biotin detection antibody, it was used as a marker substance for detection of sICAM-1 level in plasma. The sensitivity, specificity, reliability and range of detection were evaluated. The serum sICAM-1 concentrations were assayed in 32 samples by TRFIA and the results were compared with those by ELISA using correlation analysis. The serum sICAM-1 concentrations in samples of 31 healthy volunteers and 22 patients with lung disease were determined. Results The lower detection hmit was 1.32 μg/L. The inter- and intra-assay CVs were 4.24% and 6.83% , respectively. The mean recovery rate was 99.87%. The linear range of the method for sICAM-1 detection was from 2.5 to 1933 μg/L. The results measured by the TRFIA and ELISA methods were closely correlated （ R^2 = 0. 939）. There were no cross-reaction with the tumor necrosis factor （TNF） -α, interleukin （IL） -8, IL-1, IL-2 and IL-6. This method was not affected by serum factors such as hemolysis, fat and high blood serum bilirubin levels. The serum sICAM-1 concentration in the 31 health volunteers was （365.74 ± 45.03 ） μg/L. The serum sICAM-1 concentration tended to be higher in patients with inflammatory lung disease or alveolar damage. Conclusion The TRFIA of sICAM-1 has a high sensitivity and a wide detection range, making it feasible and convenient for clinical applications.

关 键 词：细胞黏着分子 荧光免疫测定 肺疾病 

分 类 号：R686[医药卫生—骨科学]