Department of Bioengineering, Faculty of Chemical and Metallurgical Engineering, Yildiz Technical University, 34210, Esenler-Istanbul, Turkey.
Sci Rep. 2017 Apr 12;7(1):852. doi: 10.1038/s41598-017-00930-1.
Toxoplasmosis is caused by T. gondii and can create serious health problems in humans and also worldwide economic harm. Because of the clinical and veterinary importance of toxoplasmosis, its timely and accurate diagnosis has a major impact on disease-fighting strategies. T. gondii surface antigen 1 (SAG1), an immunodominant-specific antigen, is often used as a diagnostic tool. Therefore, the aim of this study was the optimization of novel fluorescein isothiocyanate (FITC) labeling of the SAG1-specific IgY antibody to show the potential for immunofluorescence imaging of T. gondii in vitro. The specificity of IgY antibodies was controlled by an enzyme-linked immunosorbent assay (ELISA), and the concentration of the IgY antibody was detected using a spectrophotometer. The optimum incubation time and FITC concentration were determined with a fluorescence spectrometer. The obtained FITC-labeled IgY was used for marking T. gondii tachyzoites, which were cultured in vitro and viewed using light microscopy. The interaction of the fluorescence-labeled antibody and the T. gondii tachyzoites was examined with a fluorescence microscope. In this study, for the first time, a FITC-labeled anti-SAG1 IgY antibody was developed according to ELISA, fluorescence spectrometer, and fluorescence imaging of cell culture. In the future, the obtained FITC-labeled T. gondii tachyzoites' specific IgY antibodies may be used as diagnostic tools for the detection of T. gondii infections in different samples.
弓形虫病由刚地弓形虫引起，可在人类和全球范围内造成严重的健康问题和经济损失。由于弓形虫病的临床和兽医重要性，及时准确的诊断对疾病防控策略具有重大影响。弓形虫表面抗原 1（SAG1）是一种免疫优势特异性抗原，常被用作诊断工具。因此，本研究旨在优化新型荧光素异硫氰酸酯（FITC）标记 SAG1 特异性 IgY 抗体，以显示体外弓形虫免疫荧光成像的潜力。IgY 抗体的特异性通过酶联免疫吸附试验（ELISA）进行控制，并用分光光度计检测 IgY 抗体的浓度。用荧光分光光度计确定最佳孵育时间和 FITC 浓度。获得的 FITC 标记 IgY 用于标记体外培养的弓形虫速殖子，并通过光学显微镜观察。用荧光显微镜检查荧光标记抗体与弓形虫速殖子的相互作用。在这项研究中，首次根据 ELISA、荧光分光光度计和细胞培养荧光成像开发了 FITC 标记的抗 SAG1 IgY 抗体。将来，获得的 FITC 标记的弓形虫速殖子特异性 IgY 抗体可能被用作不同样本中弓形虫感染检测的诊断工具。
