Institute for Medical Microbiology, University Medical Center National Consulting Laboratory for Toxoplasmosis, Kreuzbergring 57, D-37075 Goettingen, Germany.
J Clin Microbiol. 2011 Jul;49(7):2419-25. doi: 10.1128/JCM.00464-11. Epub 2011 May 4.
By the separation of Toxoplasma lysate using two-dimensional gel electrophoresis and its analysis with human serum samples and mass spectrometry, the subtilisin-like protein (SUB1) was identified to be a potential marker for acute toxoplasmosis. Following expression of the C-terminal domain of SUB1 in Escherichia coli, it was tested in a line blot assay using a total of 80 human serum samples. Two computer programs based on different evaluation strategies were used for judgment of the line blot results: (i) a time-dependent method with a predefined cutoff value and (ii) a fixed-time-point method with a calculated cutoff. Thereby, SUB1 was proven to be rather reactive with specific immunoglobulin A (IgA), IgM, and IgG of patients with an acute infection. This finding makes this antigen an attractive candidate for improving diagnosis of toxoplasmosis and demonstrates that not only the selection of respective antigens but also the evaluation method chosen are important for the evaluation of new diagnostic markers.
通过二维凝胶电泳分离弓形虫裂解物，并结合人血清样本和质谱分析，鉴定枯草溶菌素样蛋白（SUB1）为急性弓形虫病的潜在标志物。在大肠杆菌中表达 SUB1 的 C 末端结构域后，使用总共 80 个人血清样本进行线状印迹分析进行了测试。使用两种基于不同评估策略的计算机程序对线状印迹结果进行判断：（i）具有预定义截止值的时间依赖性方法和（ii）具有计算截止值的固定时间点方法。因此，SUB1 被证明与急性感染患者的特异性免疫球蛋白 A（IgA）、IgM 和 IgG 反应强烈。这一发现使这种抗原成为改善弓形虫病诊断的有吸引力的候选物，并表明不仅要选择各自的抗原，还要选择所选择的评估方法，这对于评估新的诊断标志物很重要。
