ESAT-6/CFP-10 fusion protein and peptides for optimal diagnosis of Mycobacterium tuberculosis infection by ex vivo enzyme-linked immunospot assay in The Gambia
ESAT-6/CFP-10 fusion protein and peptides for optimal diagnosis of Mycobacterium tuberculosis infection by ex vivo enzyme-linked immunospot assay in The Gambia
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DOI：
10.1128/jcm.43.5.2070-2074.2005
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发表时间：
2005-05-01
影响因子：
9.4
通讯作者：
Brookes, RH
中科院分区：
文献类型：
--
作者：
Hill, PC;Jackson-Sillah, D;Brookes, RH
结核分枝杆菌抗原ESAT-6和CFP-10的重叠肽在用于γ干扰素检测的体外酶联免疫斑点（ELISPOT）测定中用于诊断结核分枝杆菌时，提供了比纯化蛋白衍生物皮肤试验更高的特异性。最近接触的结核病感染。我们评估了两种抗原的融合蛋白是否可以获得等同的结果，以及组合读数是否会在冈比亚提供更高的灵敏度。我们研究了88例痰涂阳肺结核（TB）病例的488名家庭接触者的ELISPOT检测结果。根据与结核病病例的睡眠接近程度，在暴露梯度上评估了每种测试和组合测试的阳性受试者比例。88名（18%）受试者对CFP-10肽呈阳性，148名（30%）受试者对ESAT-6肽呈阳性，161名（33%）受试者对两种肽均呈阳性，168名（34%）受试者对融合蛋白呈阳性; 188名（39%）受试者对肽呈阳性结果或对融合蛋白呈阳性结果。肽和蛋白质结果之间存在合理的一致性（kappa统计量= 0.78），无显著不一致性（P = 0.38）。在融合蛋白和组合肽斑点计数之间存在强相关性（r = 0.9），并且根据M.结核病暴露。肽库或融合蛋白阳性的受试者比例提供最大灵敏度，显著高于单独ESAT-6肽阳性的受试者比例（P = 0.007）。ESAT-6和CFP-10的融合蛋白相当于用于诊断潜伏性M的重叠肽。肺结核感染。使用肽和融合蛋白的组合提供了改进的灵敏度。
Overlapping peptides of Mycobacterium tuberculosis antigens ESAT-6 and CFP-10 offer increased specificity over the purified protein derivative skin test when they were used in an ex vivo enzyme-linked immunospot (ELISPOT) assay for gamma interferon detection for the diagnosis of M. tuberculosis infection from recent exposure. We assessed whether equivalent results could be obtained for a fusion protein of the two antigens and whether a combined readout would offer increased sensitivity in The Gambia. We studied the ELISPOT assay results for 488 household contacts of 88 sputum smear-positive tuberculosis (TB) cases. The proportions of subjects positive by each test and by the tests combined were assessed across an exposure gradient, defined according to sleeping proximity to a TB case. Eighty-eight (18%) subjects were positive for CFP-10 peptides, 148 (30%) were positive for ESAT-6 peptides, 161 (33%) were positive for both peptides, and 168 (34%) were positive for the fusion protein; 188 (39%) subjects had either a positive result for a peptide or a positive result for the fusion protein. There was reasonable agreement between the peptide and the protein results (kappa statistic = 0.78) and no significant discordance (P = 0.38). There was a strong correlation between the fusion protein and combined peptide spot counts (r = 0.9), and responses to the peptide and the proteins all increased significantly according to M. tuberculosis exposure. The proportion of subjects positive for either the pool of peptides or the fusion protein offered maximum sensitivity, being significantly higher than the proportion of subjects positive for ESAT-6 peptides alone (P = 0.007). A fusion protein of ESAT-6 and CFP-10 is equivalent to overlapping peptides for the diagnosis of latent M. tuberculosis infection. Use of a combination of peptides and fusion protein offers improved sensitivity.