目的建立基于化学发光平台的人结核感染T细胞检测方法。方法将一株γ-干扰素单抗标记在磁微粒上,另一株γ-干扰素单抗标记在吖啶酯上,然后将检测体系与已有的细胞刺激培养体系相结合。结果本研究成功建立基于化学发光平台的人结核感染T细胞检测方法。以ElISA平台检测试剂的检测结果作为参考,该方法的灵敏度为98.3%,特异性为99.2%,总体符合率达到98.8%。结论该方法具有更高的分析灵敏度(可达0.27 Pg/M l)、更宽的线性范围(1 Pg/M l~5 000 Pg/M l)、更好的重复性(批内与批间变异系数均<6.0%)及更易实现高通量检测,为临床诊断结核感染提供了有力的工具。Objective To develop the detection kit for T cell infected with Mycobacterium tuberculosis based on full-automatic chemiluminescence immune analyzer.Mehtods An anti IFN-γ Mab was coated on the surface of microparticle.Another anti IFN-γ Mab was labelled to acridinium ester.After that, the detection system was combined with the in vitro cell culture system.Results This research developed the detection kit for T cell infected with Mycobacterium tuberculosis based on full-automatic chemiluminescence immune analyzer.Compared with the testing results of the ELISA kit, the sensitivity, specificity and total matching ratio of the CLIA kit was 98.3%, 99.2% and 98.8%, respectively.Conclusion The CLIA kit has better sensitivity(0.27 pg/m L), wilder linear range(1 pg/m L ~ 5 000 pg/m L), and better repeatability(intra and inter coefficient of variation < 6.0%).It makes a high throughput detection available.It will contribute to the clinical diagnosis of Mycobacterium tuberculosis.国家高技术研究发展计划(863计划)(2011AA02A101
