Establishment of a standard 14-3-3 protein assay of cerebrospinal fluid as a diagnostic tool for Creutzfeldt-Jakob disease
Establishment of a standard 14-3-3 protein assay of cerebrospinal fluid as a diagnostic tool for Creutzfeldt-Jakob disease
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DOI：
10.1038/labinvest.2009.68
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发表时间：
2010-11-01
影响因子：
5
通讯作者：
Sata, Testutaro
中科院分区：
文献类型：
--
作者：
Satoh, Katsuya;Tobiume, Minoru;Sata, Testutaro
在脑电图记录上观察到的周期性尖波复合物和脑脊液（CSF）中14-3-3蛋白的存在都包括在世界卫生组织提供的克雅氏病（CJD）的诊断标准中;然而，由于不确定的条带，CSF中14-3-3蛋白的存在或不存在有时难以在蛋白质印迹上辨别。本研究的目的是建立一个标准的14-3-3蛋白测定法，并确定可以通过蛋白质印迹法测定的14-3-3蛋白的阈值水平。我们寻找了在蛋白质测定中测试的14-3-3蛋白质的最合适的同种型，以及对14-3-3具有亲和力的四种抗体中最敏感的抗体。我们测量了112名CJD患者和100名其他疾病患者的所有14-3-3亚型水平。我们比较了四种不同抗体的性能。我们使用LAS 3000系统对γ-亚型水平进行了半定量分析，该系统能够从蛋白质印迹上的发光产生数字图像。我们确定了进行标准化测定的14-3-3蛋白质的最合适的同种型是γ-同种型。在针对该蛋白质的四种市售抗体中，最敏感和特异性的是18647（IBL，日本）。我们报告了高重复性的检测14-3-3蛋白的γ-同种型的抗体，表明蛋白质印迹法可用于半定量分析。实验室调查（2010）90，1637-1644; doi：10.1038/labinvest.2009.68; 2010年8月9日在线发表
Periodic sharp wave complexes observed on an electroencephalographic recording and the presence of a 14-3-3 protein in the cerebrospinal fluid (CSF) are both included in the diagnostic criteria for the Creutzfeldt-Jakob disease (CJD) supplied by the World Health Organization; however, the presence or absence of the 14-3-3 protein in the CSF is sometimes difficult to discern on a western blot because of equivocal bands. The goal of this study was to establish a standard 14-3-3 protein assay and to determine the threshold level of a 14-3-3 protein that can be assayed by western blot. We searched for the most suitable isoform of the 14-3-3 protein to test for in protein assays, and the most sensitive antibody among four antibodies with an affinity for 14-3-3. We measured the levels of all 14-3-3 isoforms in 112 patients with CJD and in 100 patients with other diseases. We compared the performances of four different antibodies. We carried out a semi-quantitative analysis of gamma-isoform levels using the LAS 3000 system, which was capable of producing a digital image from the luminescence on a western blot. We determined that the most suitable isoform of the 14-3-3 protein for conducting a standardized assay was the gamma-isoform. Among the four commercially available antibodies for this protein, the most sensitive and specific was 18647 (IBL, Japan). We report the high repeatability of the detection of the 14-3-3 protein by this antibody to the gamma-isoform, showing that western blot can be used for semi-quantitative analysis. Laboratory Investigation (2010) 90, 1637-1644; doi:10.1038/labinvest.2009.68; published online 9 August 2010