Selected RD1 peptides for active tuberculosis diagnosis: Comparison of a gamma interferon whole-blood enzyme-linked immunosorbent assay and an enzyme-linked immunospot assay
Selected RD1 peptides for active tuberculosis diagnosis: Comparison of a gamma interferon whole-blood enzyme-linked immunosorbent assay and an enzyme-linked immunospot assay
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DOI：
10.1128/cdli.12.11.1311-1316.2005
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发表时间：
2005-11-01
期刊：
影响因子：
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通讯作者：
Girardi, E
中科院分区：
文献类型：
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作者：
Goletti, D;Vincenti, D;Girardi, E
我们最近建立了一种γ干扰素（ifn - γ）酶联免疫斑点试验（ELISPOT），使用选择的早期分泌抗原靶6 （ESAT-6）肽，它似乎对活动性结核病（a - tb）具有特异性。然而，ELISPOT很难实现自动化。因此，本研究的目的是确定相同选择的肽是否可以用于更适合临床实验室常规工作的技术，如全血酶联免疫吸附测定（WBE）。为此，纳入了27例A-TB患者和41例对照患者。我们的WBE使用已经描述的从ESAT-6中选择的肽和从培养滤液蛋白10中选择的三个新肽进行，并将通过ELISPOT获得的数据进行比较。使用我们选择的肽，通过WBE和ELISPOT评估，A-TB患者的ifn - γ产量显著高于对照组（P < 0.0001）。统计分析表明，WBE与ELISPOT检测结果具有良好的相关性（r = 0.80, P < 0.001）。为了证实我们的数据，我们将我们的WBE结果与QuantiFERON-TB Gold获得的结果进行了比较，QuantiFERON-TB Gold是一种基于被批准用于结核病感染诊断的RD1重叠肽的全血检测。我们观察到QuantiFERON-TB Gold的灵敏度略高于WBE(89%对81%)；然而，我们的测试提供了更好的特异性结果（90%对68%）。综上所述，基于所选RD1肽段的WBE得到的结果与ELISPOT得到的结果显著相关。此外，与QuantiFERON-TB Gold相比，我们的检测方法对a - tb诊断的特异性更高，因此它可能是临床实验室常规使用的a - tb诊断的补充工具。
We recently set up a gamma interferon (IFN-gamma) enzyme-linked immunospot assay (ELISPOT), using selected early secreted antigenic target 6 (ESAT-6) peptides, that appears specific for active tuberculosis (A-TB). However, ELISPOT is difficult to automate. Thus, the objective of this study was to determine if the same selected peptides may be used in a technique more suitable for routine work in clinical laboratories, such as whole-blood enzyme-linked immunosorbent assay (WBE). For this purpose, 27 patients with A-TB and 41 control patients were enrolled. Our WBE, using the already described selected peptides from ESAT-6 plus three new ones from culture filtrate protein 10, was performed, and data were compared with those obtained by ELISPOT. Using our selected peptides, IFN-gamma production, evaluated by both WBE and ELISPOT, was significantly higher in patients with A-TB than in controls (P < 0.0001). Statistical analysis showed a good correlation between the results obtained by WBE and ELISPOT (r = 0.80, P < 0.001). To substantiate our data, we compared our WBE results with those obtained by QuantiFERON-TB Gold, a whole-blood assay based on region of difference 1 (RD1) overlapping peptides approved for TB infection diagnosis. We observed a slightly higher sensitivity with QuantiFERON-TB Gold than with our WBE (89% versus 81%); however, our test provided a better specificity result (90% versus 68%). In conclusion, results obtained by WBE based on selected RD1 peptides significantly correlate with those generated by ELISPOT. Moreover, our assay appears more specific for A-TB diagnosis than QuantiFERON-TB Gold, and thus it may represent a complementary tool for A-TB diagnosis for routine use in clinical laboratories.