Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum.
Feasibility and diagnostic utility of antigen-specific interferon-gamma responses for rapid immunodiagnosis of tuberculosis using induced sputum.
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DOI：
10.1371/journal.pone.0010389
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发表时间：
2010-04-28
期刊：
影响因子：
3.7
通讯作者：
Dheda K
中科院分区：
文献类型：
--
作者：
Cashmore TJ;Peter JG;van Zyl-Smit RN;Semple PL;Maredza A;Meldau R;Zumla A;Nurse B;Dheda K
由于培养需要数周时间，并且难以获得代表性的生物样本，因此对涂片阴性或结核稀少型结核病（TB）的诊断存在问题。RD-1抗原特异性γ-干扰素释放试验（IGRA）是诊断支原体感染的敏感和特异的血液检测方法。肺结核感染。这种使用诱导痰细胞的快速免疫诊断试验的可行性和诊断效用尚不清楚。在101名连续招募的TB疑似患者或非TB对照中，使用标准化酶联免疫斑点（ELISPOT）测定（T-SPOT®.TB）将从诱导痰中分离的细胞与ESAT-6和CFP-10抗原共培养。使用28个样本的优化阶段之后是使用来自73名参与者的样本的验证阶段（20名患有明确或可能的TB，48名患有非TB）。尽管优化了痰液处理，但验证阶段的65/73（89%）IGRA不确定。44/73（60%）次测试由于痰液诱导相关因素[痰液诱导相关不良事件（n = 5）、痰液体积不足（n = 8）、非均质化痰液（n = 7）和进行测定的细胞数量不足（n = 24）]而失败，而20/73（27%）次测试由于T-SPOT®.TB测定相关因素[过多碎片妨碍ELISPOT孔中斑点的阅读（n = 6）、阳性对照失败（n = 11）或阴性对照中斑点计数高（n = 3）]而失败。 因此，只有8/73（11%）的可用样本可以正确分类（7例明确或可能的TB，1例非TB患者）。因此，13/20（65%）的确诊或可能的结核病例仍未得到诊断。利用诱导痰细胞，通过抗原特异性IFN-γ ELISPOT反应，可以快速免疫诊断肺结核。然而，该测试，在其目前的ELISPOT格式，是不是临床上有用的，因为大多数的测定是不确定的。
The diagnosis of smear-negative or sputum-scarce tuberculosis (TB) is problematic as culture takes several weeks and representative biological samples are difficult to obtain. RD-1 antigen-specific interferon-γ release assays (IGRAs) are sensitive and specific blood-based tests for the diagnosis of M. tuberculosis infection. The feasibility and diagnostic utility of this rapid immunodiagnostic assay, using cells from induced sputum, is unknown. Cells isolated from induced sputum were co-cultured with ESAT-6 and CFP-10 antigens using a standardized enzyme-linked immunospot (ELISPOT) assay (T-SPOT®.TB) in 101 consecutively recruited TB suspects or non-TB controls. An optimization phase using 28 samples was followed by a validation phase using samples from 73 participants (20 with definite or probable TB, and 48 with non-TB). Despite optimization of sputum processing 65/73 (89%) of the IGRAs in the validation phase were inconclusive. 44/73 (60%) tests failed due to sputum induction-related factors [sputum induction-related adverse events (n = 5), inadequate sputum volume (n = 8), non-homogenisable sputum (n = 7), and insufficient numbers of cells to perform the assay (n = 24)], whilst 20/73 (27%) tests failed due T-SPOT®.TB assay-related factors [excessive debris precluding reading of spots in the ELISPOT well (n = 6), failure of the positive control (n = 11), or high spot count in the negative control (n = 3)]. Only 8/73 (11%) of the available samples could therefore be correctly categorized (7 definite or probable TB, and 1 non-TB patient). Thus, 13/20 (65%) of the definite or probable TB cases remained undiagnosed. Rapid immunodiagnosis of pulmonary TB by antigen-specific IFN-γ ELISPOT responses, using cells from induced sputum, is possible. However, the test, in its current ELISPOT format, is not clinically useful because the majority of the assays are inconclusive.
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影响因子：
3.3
作者：
Dheda, K;Udwadia, ZF;Rook, GAW
通讯作者：
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DOI：
10.1136/ebm.13.1.14
发表时间：
2008-02-01
期刊：
Evidence-based medicine
影响因子：
--
作者：
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DOI：
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发表时间：
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影响因子：
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DOI：
10.1164/ajrccm.162.6.2003125
发表时间：
2000-12-01
影响因子：
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作者：
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通讯作者：
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