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目的：建立及评价使用磁性纳米微球作为固相载体的人酌干扰素（Interferon-gamma，IFN-gamma）双抗体夹心酶联免疫吸附实验（Enzyme-linked immunosorbent assay，ELISA）检测方法。方法：以杂化细乳液合成法制备磁性纳米微球，将其作为免疫检测的固相载体。将磁性微球与IFN-酌抗体进行偶联,建立基于磁性微球的ELISA 检测方法，检测人IFN-gamma，绘制IFN-gamma标准曲线并进行方法学评价。结果：获得包被有人IFN-gamma抗体的免疫微球, 抗体偶联率为54.5 %。用它建立IFN-gamma的双抗体夹心的ELISA 检测方法，检测范围为0-1000 pg/mL，相关系数为0.9996，灵敏度23.2 pg/mL，功能灵敏度0 pg/mL，批内和批间变异系数（Coefficients of Variance，CVs）＜8 %，检测总共需要2 小时。结论：成功制备了IFN-酌免疫微球并建立了定量检测人IFN-gamma的双抗体夹心磁珠酶联免疫方法。

英文摘要:

Objective:To establish and evaluate a double Interferon-gamma (IFN-gamma) antibody sandwich Enzyme-linked immunosorbent assay（ELISA）which was based on immunomagnetic beads for human IFN-gamma detection.Methods:Magnetic nanoparticle were prepared through a hybrid miniemulsion method and coupled with hunman IFN-Gamma antibody as solid phase. IFN-Gamma Antibody was coupled to magnetic nanoparticle. ELISA based on immunomagnetic beads for human IFN-Gamma detection was established and evaluated. Then the standard curve was drawn and assay performance characteristics of the new method was evaluated.Results:Immunoparticles coated human IFN-gamma antibody were obtained with a coupling efficiency of 54.5 %. Immunoparticles were used to establish the double IFN-gamma antibody sandwich system with detection range of 0-1000 pg/mL, coefficiency of correlation of 0.9996, sensitivity of 23.2 pg/mL, functional sensitivity of 0 pg/mL, and intra- and inter-assay coefficients of variation (CVs)＜8 %. The total test time was 2 hours.Conclusion:A sandwich ELISA was successfully established for quantitative IFN-gamma detection based on immunomagnetic beads.

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