Specific T-cell epitopes for immunoassay-based diagnosis of Mycobacterium tuberculosis infection
Specific T-cell epitopes for immunoassay-based diagnosis of Mycobacterium tuberculosis infection
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DOI：
10.1128/jcm.42.6.2379-2387.2004
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发表时间：
2004-06-01
影响因子：
9.4
通讯作者：
Andersen, P
中科院分区：
文献类型：
--
作者：
Brock, I;Weldingh, K;Andersen, P
目前用于结核感染的免疫学检测方法，即结核菌素皮肤试验，特异性低。因此，迫切需要结核分枝杆菌特异性抗原来替代纯化的蛋白衍生物。我们对最近发现的四种抗原（Rv 2653、Rv 2654、Rv 3873和Rv 3878）的诊断潜力进行了严格评估，这些抗原来自牛分支杆菌卡介苗（BCG）疫苗株以及最常见的非结核分支杆菌缺乏的基因组区域。这些分子中潜在表位的精细特异性通过来自M的外周血单个核细胞的T细胞应答的敏感性测试来评估。牛BCG疫苗接种的健康个体与合成的重叠肽。四种分子中的三种含有具有显著特异性问题的区域（Rv 2653、Rv 3873和Rv 3878）。我们从四种蛋白质中选择并组合了不被M识别的特定肽段。接种BCG疫苗的个体。这些肽段用从显微镜或培养证实的结核病患者和健康的结核分枝杆菌获得的外周血单核细胞进行测试。牛BCG接种对照。该分析中最有希望的延伸段的组合显示出与两种众所周知的M.结核特异性蛋白ESAT-6和CFP-10（分别为75%和66%）。ESAT-6、CFP-10和新型特异性肽段的组合给出了84%的总体灵敏度和97%的特异性。在使用新实验组的验证实验中，对于肽的组合获得的灵敏度为57%，对于肽、ESAT-6和CFP-10的组合获得的灵敏度为90%。该组合的特异性为95%。
The currently used method for immunological detection of tuberculosis infection, the tuberculin skin test, has low specificity. Antigens specific for Mycobacterium tuberculosis to replace purified protein derivative are therefore urgently needed. We have performed a rigorous assessment of the diagnostic potential of four recently identified antigens (Rv2653, Rv2654, Rv3873, and Rv3878) from genomic regions that are lacking from the Mycobacterium bovis bacillus Calmette-Guerin (BCG) vaccine strains as well as from the most common nontuberculous mycobacteria. The fine specificity of potential epitopes in these molecules was evaluated by sensitive testing of the T-cell responses of peripheral blood mononuclear cells derived from M. bovis BCG-vaccinated healthy individuals to synthesized overlapping peptides. Three of the four molecules contained regions with significant specificity problems (Rv2653, Rv3873, and Rv3878). We selected and combined the specific peptide stretches from the four proteins not recognized by M. bovis BCG-vaccinated individuals. These peptide stretches were tested with peripheral blood mononuclear cells obtained from patients with microscopy or culture-confirmed tuberculosis and from healthy M. bovis BCG-vaccinated controls. The combination of the most promising stretches from this analysis showed a sensitivity level (57%) comparable to the level found with the two well-known M. tuberculosis-specific proteins ESAT-6 and CFP-10 (75 and 66%, respectively). The combination of ESAT-6, CFP-10, and the novel specific peptide stretches gave an overall sensitivity of 84% at a specificity of 97%. In a validation experiment with new experimental groups, the sensitivities obtained were 57% for the combination of peptides and 90% for the combination of the peptides, ESAT-6, and CFP-10. This combination gave a specificity of 95%.