四种结核抗原的诊断效能分析与建立结核抗原诊断效能的评价体系

目的通过对4种结核分枝杆菌抗原的诊断效能分析,初步建立其诊断效能评价体系,为结核病免疫学诊断筛选特异、有效抗原提供根据。方法采用酶联免疫吸附试验(ELISA)和受试者工作特征曲线法(Receiver Operating Characteristic,ROC)建立结核抗原诊断效能评价体系。然后,对4种重组结核抗原的诊断效能进行分析,得出各种抗原组合的检测阳性率,进而评价得出最佳的抗原组合方式。结果该结核抗原诊断效能的评价体系包括最佳ELISA反应体系、ROC曲线的建立、确定各种抗原最佳临界值(Cut-off value)、最佳抗原组合方式的确定。ELISA检测体系的中CFP10、38kDa、MPT64和Ag85B的最佳蛋白包被浓度分别为0.5μg/mL、2μg/mL、2μg/mL和1μg/mL,最佳血清稀释度分别为1∶250、1∶225、1∶225和1∶225;对这4种结核抗原及其不同组合的ELISA检测诊断效能进行了评价,以组合抗原的诊断效能较好,最佳组合为38kDa+CFP10+MPT64,阳性率为64.7%。结论成功建立了结核抗原诊断效能评价体系。结果显示ELISA检测中以多组分抗原组合为宜。

The diagnostic effect analysis of 4 Mycobacterium tuberculosis antigens and the establishment of one diagnosis effect evaluation system

With the diagnosis effect analysis of 4 recombinant tuberculosis protein antigens, the evaluation system was established by the Enzyme-Linked Immunosorbnent Assay （ELISA） and the Receiver Operating Characteristic curve （ROC）, by which the diagnostic effect of the 4 antigens were analyzed . The positive rate of every possible combination of the 4 antigens was calculated, then the best combination was evaluated. This evaluation systems including estsblishment of optimal ELISA conditions,ROC curves optimal Cut-off value of each antigen and optimal combinatorial way; The optimal concentration of the recombinant proteins CFP10, 38kDa, MPT64 and Ag85B used in ELISA were 0.5μg/mL, 2μg/mL, 2 μg/mL and 1μg/mL and the the best serum dilution were 1 ： 250,1 ： 225,1 ： 225 and 1 ： 225 respectively; the effect of combination was much better than any single antigen, 38kDa＋CFP10＋MPT64 was the best combination of all, of which the positive rate was 64.7%. The evaluation system of diagnostic effect for M. tuberculosis antigens has been established preliminarily. According to the results of this study, it would be better to choose multi-antigen combination in ELISA detection.