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作 者:袁仕善[1] 易新元[1] 曾宪芳[1] 王敏[1] 张顺科[1] Larry Mc Reynolds
机构地区:[1]中南大学湘雅医学院血吸虫病研究室,长沙市410078 [2]MolecularParasitologyGroup,NewEnglandBiolabs
出 处:《中国热带医学》2002年第4期441-443,共3页China Tropical Medicine
基 金:湖南省科技厅资助项目 (0 0jzy2 1 1 5);WHO/TDR资助项目 (980 268)
摘 要:目的 筛选日本血吸虫成虫 67kD抗原的模拟表位 ,对其免疫学活性予以鉴定。方法 用粗提日本血吸虫成虫 67kD抗原的抗体IgG作配体筛选噬菌体 12肽库 ,随机挑取三轮筛选后的噬菌体克隆 ,Dot -ELISA检测其特异性 :用混和阳性噬菌体克隆免疫小鼠 3次 ,攻击感染后 45d剖杀 ,计数虫荷。结果 经三轮筛选 ,特异噬菌体得到富集 ,挑取的 11个噬菌体克隆经Dot-ELISA鉴定均与 67kD抗原免疫血清呈阳性反应。混合噬菌体克隆的免疫血清可识别 67kD抗原 ,并具有一定的抗血吸虫的免疫保护效果。结论 利用噬菌体随机肽库技术获得了日本血吸虫成虫67kD抗原的模拟表位 。Objective To analyse epitopes of Schistosoma japonicum(SJ) adult worm 67kD antigen and explore its immunologic activities.Methods Phage random peptide library was immunoscreened with purified IgG from sera of mice immunized with Sj adult worm 67kD antigen.Positive clones were detected by Dot-ELISA,which were obtained after three rounds of biopanning.Mice were immunized with mixed positive phage clones for three times,All mice were perfused 45 days after challenge with Sj cercariae.The worms were counted.Result The specific phages binding to IgG were enriched after three rounds of biopanning.Eleven clones detected by Dot-ELISA bound to the specific IgG of sera immunized with 67kD antigen.Immunoprotection was imduced by immunization of mixed positive phage clones.Conclusion The specific epitopes mimicking 67kD antigen were obtained by immunoscreening phage random peptide library and the specific epitopes possess good immunogenicity.
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