基于磁颗粒化学发光免疫分析方法检测鳞状细胞癌抗原∗

摘要

目的：建立一种快速、灵敏的化学发光免疫分析方法检测人血清中鳞状细胞癌抗原(SCCA)水平。方法使用异鲁米诺(ABEI)和异硫氰酸荧光素(FITC)分别标记 SCCA 的单克隆抗体,与待测 SCCA 抗原通过夹心法免疫反应形成抗原抗体复合物,采用包被有 FITC 的磁颗粒作为固相分离载体,加入底物之后检测发光强度。结果本方法线性范围达到22 ng/mL,灵敏度为0.025 ng/mL,批内变异系数(CV)和批间 CV 分别小于6%和7%。与现有的 SCCA 检测方法进行比对,相关系数为0.9901。结论基于磁颗粒化学发光免疫分析检测 SCCA 的方法性能稳定、可靠,可用于定量检测人血清中 SCCA 浓度。%Objective To establish a rapid and sensitive chemiluminescence immunoassay for detecting squamous cell carcinoma antigen (SCCA)in human serum.Methods Fluorescein isothiocyanate (FITC)and N-(aminobutyl)-N-(ethylisoluminol)(ABEI) were used to label two different monoclonal antibodies of anti-SCCA.SCCA in serum combined with labeled antibodies and formed a sandwiched immunoreaction.After adding the substrate solution,the relative light unit of ABEI was measured.Magnetic particles coated with anti-FITC antibody were used as solid separation carrier.Results The results demonstrated that the method was linear to 22 ng/mL with a detection limit of 0.025 ng/mL.The coefficient of variation (CV)was less than 6% and 7% for intra-assay and inter-assay precision,respectively.Compared with the commercial kit,the proposed method showed a correlation of 0.990 1.Conclu-sion chemiluminescence immunoassay based on magnetic particles displays acceptable performance for quantification of SCCA and is appropriate for use in clinical diagnosis.