华支睾吸虫模拟抗原表位的筛选和鉴定Ξ.pdfVIP

672 　　　　　　　　　　　　　　　CHINA TROPICAL MEDICINE Vol7 No5 May 2007 　中国热带医学 2007 年第 7 卷第 5 期 [ 论 　　著] 华支睾吸虫模拟抗原表位的筛选和鉴定 余杨 ,刘宜升 ,付琳琳 ,杜文平 ,郑葵阳 摘要 : 目的　从噬菌体 12 肽库中筛选华支睾吸虫模拟抗原表位 。　方法 　应用噬菌体随机 12 肽库亲和筛选华 支睾吸虫病患者混合血清 IgG ,经过 3 轮的淘筛 ,随机挑取蓝色噬菌斑进行扩增 , 以 ELISA 检测其免疫活性 ,挑选免疫 活性较高的克隆进行序列分析及免疫学鉴定 。　结果 　8 个阳性克隆测得 6 个 DNA 序列 ,Western - bloting 显示 6 个 单克隆噬菌体均可与华支睾吸虫患者血清反应 ,斑点免疫金银染色法有 3 个克隆可被华支睾吸虫患者血清识别 。　 结论 　应用噬菌体肽库筛选技术可以获得华支睾吸虫模拟抗原表位 。 关键词 :华支睾吸虫 ;噬菌体肽库 ;抗原模拟表位 ( ) 中图分类号 :R53223 　　文献标识码 :A 　　文章编号 :1009 - 9727 2007 5 - 672 - 03 Screening and identification of mimic antigen epitopes of Clonorchis sinensis . YU Yang , LIU Yi - sheng , FU Lin - lin , et al . (Department of Pathogen and Biology , Xuzhou Medical College , Xuzhou 221002 , Jiangsu , P. R. China) Screening and identification of mimic antigen epitopes of Clonorchis sinensis. YAbstract :Objective 　To screen antigenic epi tope of Chonorchis sinensis from phage 12 - mer peptide library. 　Methods 　Three rounds of affinity selection with immunoglobulin Gfrom pooled sera of Clonorchis sinensis patients were carried out by using a phage 12 - mer peptide library. Blue plaques were picked out randomly and amplified ,then clones with high immunocompetence were detected by sequential analysis by ELISA and im munology identification . 　Results 　The sequences of the 6 positive phage clones were determined. The result of western - blotting indicates that 6 clones could all bind to sera from patients infected with Clonorchis sinensis . Dot IGSS showed 3 clones can be dis cerned