结核病不同抗原酶联免疫斑点检测的影响因素分析

The Effect of Different Antigens on Enzyme - linked Immunospot Assay for Tuberculosis

目的比较不同MTB抗原的酶联免疫斑点检测技术（ELISPOT）与各种影响因素的关系。方法对结核组132例和对照组44例观察对象应用PPD、克隆表达的早期分泌抗原靶6000（ESAT-6）和ESAT．6／培养滤液蛋白10000（CFP．10）融合蛋白（E／C）进行酶联免疫斑点检测（ELISPOT），检测外周血单个核细胞（PBMC）中经3种抗原刺激后斑点形成细胞（SFC）的数量。计量资料用中位数（四分位间距）表示。计量资料的比较采用Mann—Whimeyu检验，计数资料的比较采用x^2检验。结果E／C—ELISPOT在治疗时间〉15d病例形成的SFC数量少于治疗时间≤15d形成的SFC数量。E/C—ELISPOT形成SFC数量与其他因素包括年龄、性别、病史等因素影响。PPD—ELISPOT和ESAT-6-ELISPOT形成SFC数量不受年龄、性别、病史、抗结核治疗时间等因素影响。3种抗原ELISPOT的敏感性不受年龄、性别、病史、抗结核治疗时间等因素影响。结论ESAT6／CFP-10融合蛋白-ELISPOT形成的斑点数受治疗时间影响，可能成为监测抗结核治疗疗效的一项指标。

Objective To compare the effect of differentMycobacterium tuberculosis antigens on enzyme-linked immunospot assay （ELISPOT）. Methods 132 tuberculosis patients and 90 control cases were recruited to the research. The peripheral blood mononuclear cells （PBMC） of all partici- pants were co-cultured with the following antigens： purified protein derivative （ PPD）, early secretory antigenic target-6 （ESAT-6） and ESAT-6/culture filtrate protein-10 fusion protein （ ESAT-6/CFP-10, E/C）. Spot forming cells （SFC） were enumerated by ELISPOT. Tuberculin skin test was performed to all control participants. Results SFC count detected by ESAT-6 / CFP-10 fusion protein ELISPOT were correlated with the duration of therapy. Conclusions ESAT-6 / CFP 10 fusion protein-ELISPOT may act as a marker of antituberculous chemotherapy.