Department of Biomedical Engineering, Medical School, Shenzhen University, Guangdong, PR China.
Clin Chim Acta. 2011 Aug 17;412(17-18):1572-7. doi: 10.1016/j.cca.2011.05.005. Epub 2011 May 12.
Because squamous cell carcinoma antigen (SCCa) quantification has demonstrated strong clinical potential, we describe a rapid and highly sensitive magnetic particle-based chemiluminescence immunoassay (CLIA) technique for assaying SCCa in serum.
Fluorescein isothiocyanate (FITC) and N-(aminobutyl)-N-(ethylisoluminol) (ABEI) were used to label 2 different monoclonal antibodies of anti-SCCa. Both of the labeled antibodies combined with SCCa to form a sandwiched immunoreaction that was monitored by chemiluminescence (CL) detection. The magnetic particles (MPs) that were coated with anti-FITC antibody served as both the solid phase and the separator. The relevant variables involved in the CLIA signals were optimized and the parameters of the proposed method were evaluated.
The method was linear to 20 ng/ml SCCa with a detection limit of 0.02 ng/ml. The intra-assay imprecision results [mean (CV)] were 1.12 ng/ml (3.81%), 2.58 ng/ml (2.53%) and [6.56 ng/ml (2.24%)]; the inter-assay imprecision results were [1.18 ng/ml (5.26%)], [2.49 ng/ml (4.75%)] and [6.61 ng/ml (4.29%)]. The average recoveries were between 97% and 104%. The relationship between the concentration of diluted SCCa and the dilution ratios gave a linear correlation coefficient of 0.9995. A correlation analysis against an established automated assay generated a slope of 0.9929 and an intercept of 0.0039 ng/ml (r=0.9964).
The proposed method demonstrates an acceptable performance for quantifying serum SCCa and is suitable for the fabrication of a commercial kit with application in the automated CL analyzer.
由于鳞状细胞癌抗原 (SCCa) 的定量分析具有很强的临床应用潜力，我们描述了一种快速、高灵敏度的基于磁性粒子的化学发光免疫分析 (CLIA) 技术，用于检测血清中的 SCCa。
使用荧光素异硫氰酸酯 (FITC) 和 N-(氨丁基)-N-(乙基异鲁米诺) (ABEI) 标记抗 SCCa 的 2 种单克隆抗体。两种标记的抗体与 SCCa 结合形成夹心免疫反应，通过化学发光 (CL) 检测进行监测。涂有抗 FITC 抗体的磁性粒子 (MPs) 既作为固相又作为分离剂。对 CLIA 信号涉及的相关变量进行了优化，并对该方法的参数进行了评估。
该方法在 20ng/ml SCCa 范围内呈线性，检测限为 0.02ng/ml。批内精密度试验结果[均值 (CV)]分别为 1.12ng/ml (3.81%)、2.58ng/ml (2.53%)和[6.56ng/ml (2.24%)]；批间精密度试验结果分别为[1.18ng/ml (5.26%)]、[2.49ng/ml (4.75%)]和[6.61ng/ml (4.29%)]。平均回收率在 97%到 104%之间。稀释 SCCa 浓度与稀释倍数之间的关系呈线性相关，相关系数为 0.9995。与已建立的自动化检测方法的相关性分析产生了 0.9929 的斜率和 0.0039ng/ml 的截距 (r=0.9964)。
该方法在定量检测血清 SCCa 方面具有良好的性能，适用于制作商业试剂盒，并可应用于自动化 CL 分析仪。
