Sequential recognition of antigenic markers of Toxoplasma gondii tachyzoite by pooled sera of mice with experimental toxoplasmosis
Sequential recognition of antigenic markers of Toxoplasma gondii tachyzoite by pooled sera of mice with experimental toxoplasmosis
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DOI：
10.1007/s00436-010-2046-0
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发表时间：
2011-01-01
影响因子：
2
通讯作者：
Abdel-Razek, Mona Gamal
中科院分区：
文献类型：
--
作者：
Habib, Fayza Sayed Mohamed;Ali, Nehad Mahmoud;Abdel-Razek, Mona Gamal
对母体弓形虫病的准确诊断可以提高医疗的成功率，防止先天性传播。目前的诊断方法有许多局限性，它们很难区分最近和潜伏感染。本工作进行了记录的抗原标记物的弓形虫速殖子全提取物和糖基肌醇磷脂（GIPL）富集的馏分特异性IgG和IgM的顺序识别，分别通过免疫印迹分析的抗原对每日合并的血清样品与实验诱导的最近和潜伏的弓形虫病小鼠。IgG亲合力免疫印迹通过使用用6 M尿素溶液作为抗原-抗体破坏剂的洗涤来测试。最近感染的小鼠合并血清中，10 kDa的条带仅与低亲合力IgG反应。39 kDa的条带是感染的良好标志物，在近期感染中与低亲和力IgG反应，在潜伏感染中与高亲和力IgG反应。15，23，30，60，66，和97 kDa的条带在感染的两个阶段与可变的亲合力反应。通过十二烷基硫酸钠-聚丙烯酰胺凝胶电泳检测到两条抗原带，即14-和30-kDa带。14-kDa的带被IgM识别在最近感染的小鼠的合并血清样品中，而30-kDa的带被识别的最近和潜伏期的感染的血清样品。这项研究强调了亲合力免疫印迹法在区分近期和潜伏实验性弓形虫病中的价值。进一步的研究必须在人体上进行，以评估所使用的技术的价值。
Accurate diagnosis of maternal toxoplasmosis can enhance the success of medical treatment and prevent congenital transmission. The current diagnostic methods have many limits, and they poorly differentiate between recent and latent infections. The present work was conducted to record the sequential recognition of antigenic markers of both Toxoplasma tachyzoites whole extract and glycosylinositolphospholipids (GIPLs)-enriched fraction by specific IgG and IgM, respectively, by immunoblotting analysis of the antigens against daily pooled serum samples from mice with experimentally induced recent and latent toxoplasmosis. IgG avidity immunoblotting was tested by using a wash with 6 M urea solution as antigen-antibody disrupting agent. Band of 10 kDa reacted exclusively with low-avidity IgG in pooled sera of mice with recent infection. Band of 39 kDa was a good marker for the infection; reacting with both low-avidity IgG in recent infection and with high-avidity IgG in latent one. Bands of 15, 23, 30, 60, 66, and 97 kDa reacted with variable avidity in both phases of infection. Two antigenic bands were detected by sodium dodecyl sulfate-polyacrylamide gel electrophoresis of the GIPLs-enriched fraction of tachyzoite, the 14- and 30-kDa band. The 14-kDa band was recognized by IgM in pooled serum samples of recently infected mice only, while the 30-kDa band was recognized by serum samples of both recent and latent phases of infections. The study highlights the value of avidity immunoblotting assay to discriminate between recent and latent experimental toxoplasmosis. Further study must be carried on human to evaluate the values of the used technique.