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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。
OBJECTIVE: To screen short peptide fragments that can specifically bind to the serum of patients with chronic schistosomiasis from phage random peptide library. Methods Serum immunoglobulin (Ig) of patients with chronic schistosomiasis was used as a ligand to immunoscreen a random 7-peptide library expressed as fusion protein in coat protein III of filamentous bacteriophage M13. After three rounds of panning, the phage clones were randomly picked up and tested for specificity by ELISA. The value of dot ELISA was analyzed for the diagnosis of schistosomiasis. Results After 3 rounds of panning, the specific binding phage enrichment increased nearly 100-fold. Thirty-six phage clones randomly selected after the third round of screening by ELISA were specifically reactive with serum Ig in patients with chronic schistosomiasis. Two of the clones with the highest A value had the potential value of diagnosing schistosomiasis . Conclusion The phage random peptide library technology can be applied in the analysis and identification of schistosome antigen epitopes, and then provide the basis for obtaining the diagnostic reagent of subunit epitope level and short peptide vaccine against schistosomiasis.