Manoutcharian K, Sotelo J, Garcia E, Cano A, Gevorkian G
Instituto de Investigaciones Biomedicas, Universidad Nacional Autonoma de Mexico, Mexico.
Clin Immunol. 1999 Apr;91(1):117-21. doi: 10.1006/clim.1998.4669.
To identify epitopes for antibodies present in cerebrospinal fluid (CSF) samples from two patients with confirmed neurocysticercosis, we used a phage peptide library that displayed random dodecapeptides as a fusion to the minor coat protein (pIII) of phage M13. To increase the specificity of selection, plates coated with anti-human Fc antibody were used in five rounds of biopanning. The DNA inserts of 30 selected clones were determined and the deduced amino acid sequences were analyzed. Sequence similarities between affinity-selected clones and three known T. solium and T. crassiceps proteins were encountered. Two phage clones have been shown to react in enzyme-linked immunosorbent assay with CSF samples from other patients with neurocysticercosis, and no reaction was observed with CSF samples from patients with other neurological disorders used as negative controls. Our results suggest that affinity selection of peptides from phage libraries using CSF samples from patients is an attractive tool for the development of novel reagents for a simple and sensitive immunodiagnostic test and for understanding the molecular mechanisms participating in the pathogenesis of neurocysticercosis.
为了鉴定来自两名确诊为神经囊尾蚴病患者的脑脊液（CSF）样本中存在的抗体的表位，我们使用了一个噬菌体肽库，该库展示了作为噬菌体M13次要外壳蛋白（pIII）融合体的随机十二肽。为了提高筛选的特异性，在五轮生物淘选中使用了包被抗人Fc抗体的平板。确定了30个选定克隆的DNA插入片段，并对推导的氨基酸序列进行了分析。发现亲和选择的克隆与三种已知的猪带绦虫和肥胖带绦虫蛋白质之间存在序列相似性。已证明两个噬菌体克隆在酶联免疫吸附试验中与其他神经囊尾蚴病患者的脑脊液样本发生反应，而用作阴性对照的其他神经系统疾病患者的脑脊液样本未观察到反应。我们的结果表明，使用患者的脑脊液样本从噬菌体库中亲和选择肽是开发用于简单、灵敏免疫诊断测试的新型试剂以及理解参与神经囊尾蚴病发病机制的分子机制的有吸引力的工具。
