流动注射协同增强化学发光免疫分析法的建立与评价

Establishment and Evaluation of an Approach of Flow Injection Immunoassay with a Synergistic Enhanced Chemiluminescent System

以兔IgG为模型 ,NaTPB与PPP协同增强的Luminol H2 O2 HRP混合液为发光体系 ,HRP标记抗体 ,建立了灵敏度高、特异性强、重现性好的流动注射协同增强化学发光免疫分析法。结果表明 :在 2～ 6 0 μg L范围内兔IgG量与发光强度有良好的线性关系 ,相关系数r=0 .994 1 (P <0 .0 1 ) ;绝对检测限为 0 .6 5fmol;方法精密度为 4 .72 %～ 9.31 % ;回收率为 92 .5 0 %～ 99.4 0 %。免疫柱可反复使用 2 0 0次以上。

A new technique of flow injection immunoassay with a synergistic enhanced effect of sodium tetraphenylborate and para-phenylphenol on the luminol-H 2O 2-horseradish peroxidase chemiluminescent system was applied to the determination of rabbit immunoglobulin (IgG), Satisfactory results were obtained in the dynamic range 2～60 μg/L of rabbit IgG with a detection limit of 0.65 fmol and a precision (RSD) of 4.7%～9.3%. Recovery of the assay was 92.3%～99.4% for rabbit IgG. The lifetime of the immuo-column was long enough, one column was applied for more than 200 determinations in one month without significant loss of activity.The results of rabbit IgG determination accorded with those of radio immunoassay. The approach put together the techniques of chemiluminescent analysis, flow injection analysis and immunoassay, concentrated all their advantages of the high sensitivity, high precision and high selectivity.