日本血吸虫病患者血清对噬菌体随机7肽库的免疫筛选  

Immunoscreening of phage random peptide library with sera from patients with schistosomiasis japonica

作　　者：舒新华[1] 易新元[1] 曾宪芳[1] 

机构地区：[1]湖南医科大学寄生虫学教研室,湖南长沙410078

出　　处：《细胞与分子免疫学杂志》2000年第2期109-112,共4页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家九五医学科技攻关项目!资助 ;No.96 06 04 04

摘　　要：目的从噬菌体随机多肽文库中 ,筛选出能与慢性血吸虫病患者血清特异性结合的短肽分子。方法采用慢性血吸虫病患者血清免疫球蛋白(Ig)作为配基 ,免疫筛选以融合蛋白形式在丝状噬菌体M13外壳蛋白III表达的随机7肽文库。按吸附 -洗脱 -扩增的淘筛过程 ,经3轮免疫淘筛后 ,随机挑取噬菌体克隆用ELISA检测其特异性 ,并用斑点ELISA分析其诊断血吸虫病的价值。结果经3轮免疫淘筛后 ,特异性结合的噬菌体富集增加近100倍。用ELISA检测第3轮筛选后随机挑取的30个噬菌体克隆 ,有26个克隆能与慢性血吸虫病患者的血清Ig产生特异性反应 ,其中A值最高的2个克隆具有诊断血吸虫病的潜在价值。结论噬菌体随机肽库技术 ,可应用于分析、鉴定血吸虫抗原表位 ,继而为获得亚单位表位水平的诊断试剂和抗血吸虫病的短肽疫苗提供依据。Aim To obtain the short peptides through screening phage random peptide library with sera from schistosomiasis patients. Methods Specific Ig was purified from sera of patients with chronic schistosomiasis and used to immunoscreen a phage random peptide library of 7 anmino acid residues displayed as a fusion protein on shell protein III of filamentous phage M13. The positive clones were obtained through three rounds of biopanning; the reactivity of each clone bound to specific Ig was examined by ELISA. The specific clones were used to examine sera from schistosomiasis patients by Dot ELISA. Results The eluted phages were enriched nearly to 100 fold through three rounds of biopanning. In 30 phage clones selected randonly from the third round biopanning, 26 clones of them could bind to the specific Ig. Two clones with highest A595 value in positive clones had a potential role in immunodiagnosis of schistosomiasis. Conclusion Schistosome specific antigenic epitopes can be identified by the technique of immunoscreening phage random peptide library using infected patient sera and these epitopes pay the foundation for preparing immunodiagnostic reagent or vaccine.

关 键 词：日本血吸虫病 免疫筛选 噬菌体 随机多肽文库 

分 类 号：R532.210.3[医药卫生—内科学]