High sensitivity of an ELISA kit for detection of the gamma-isoform of 14-3-3 proteins: usefulness in laboratory diagnosis of human prion disease.
High sensitivity of an ELISA kit for detection of the gamma-isoform of 14-3-3 proteins: usefulness in laboratory diagnosis of human prion disease.
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DOI：
10.1186/1471-2377-11-120
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发表时间：
2011-10-04
期刊：
影响因子：
2.6
通讯作者：
Nishida N
中科院分区：
文献类型：
--
作者：
Matsui Y;Satoh K;Miyazaki T;Shirabe S;Atarashi R;Mutsukura K;Satoh A;Kataoka Y;Nishida N
14-3-3 蛋白的 γ 异构体 (14-3-3 gamma) 在神经元中表达，可能是神经元损伤的特异性标志物。据报道，这种蛋白质是一种可检测的生物标志物，特别是通过蛋白质印迹 (WB) 或酶联免疫吸附测定 (ELISA) 在克雅氏病 (CJD) 患者的脑脊液 (CSF) 中发现。 14-3-3 γ 的蛋白质印迹法不敏感，并且报告的数据在出版物之间存在冲突。没有针对 14-3-3 gamma 的特异性 ELISA。本研究对克雅氏病患者（n = 114 名散发性克雅氏病患者、7 名遗传性克雅氏病患者和 3 名医源性克雅氏病患者）和 99 名患有其他神经退行性疾病的患者进行了检查。通过蛋白质印迹分析获得的脑脊液样本的 14-3-3 γ，并通过 ELISA 分析总 tau 蛋白。我们评估了新开发的夹心 ELISA 对 14-3-3 γ 的敏感性和特异性。 14-3-3 γ ELISA 的截止值> 1, 683 AU/ml；敏感性为 95.2%，特异性为 72.7%。这种特异性与总 tau 蛋白 ELISA 相同。 7 例克雅氏病 WB 呈阴性，但 14-3-3 γ ELISA 呈阳性，表明 ELISA 更敏感。结合 14-3-3γ ELISA 和弥散加权 MR 成像 (DWI-MRI) 可以诊断所有 21 例早期克雅氏病。 14-3-3 γ ELISA 比传统 WB 更灵敏，可用于 CJD 的实验室诊断，类似于 tau 蛋白的 ELISA。使用 DWI-MRI 和这些脑脊液 ELISA 测试，即使在疾病的早期阶段也可以诊断克雅氏病。
The gamma-isoform of the 14-3-3 protein (14-3-3 gamma) is expressed in neurons, and could be a specific marker for neuronal damage. This protein has been reported as a detectable biomarker, especially in the cerebrospinal fluid (CSF) of Creutzfeldt-Jakob disease (CJD) patients by Western blotting (WB) or enzyme-linked immunosorbent assays (ELISAs). Western blotting for 14-3-3 gamma is not sensitive, and the reported data are conflicting among publications. An ELISA specific for 14-3-3 gamma is not available. CJD patients (n = 114 sporadic CJD patients, 7 genetic CJD, and 3 iatrogenic CJD) and 99 patients with other neurodegenerative diseases were examined in this study. The CSF samples obtained were analyzed by Western blotting for 14-3-3 gamma, and by ELISA for total tau protein. We evaluated the sensitivity and specificity of the newly developed sandwich ELISA for 14-3-3 gamma. The cut-off value of the 14-3-3 gamma ELISA was > 1, 683 AU/ml; and sensitivity was 95.2%, with 72.7% specificity. This specificity was the same for the total tau protein ELISA. Seven CJD cases were negative by WB but positive using the 14-3-3 gamma ELISA, indicating that the ELISA is more sensitive. All 21 cases of early stage CJD could be diagnosed using a combination of the 14-3-3γ ELISA and diffusion weighted MR imaging (DWI-MRI). The 14-3-3 gamma ELISA was more sensitive than conventional WB, and was useful for laboratory diagnosis of CJD, similar to the ELISA for the tau protein. Using DWI-MRI and these ELISA tests on CSF, diagnosis of CJD will be possible even at early stages of the disease.
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