Qian Min, Bai Yan-jun, Zhang Ping, Yan Zi-zhu, Wu Zi-rong
School of Life Science, East China Normal University, Shanghai 200062.
Zhongguo Ji Sheng Chong Xue Yu Ji Sheng Chong Bing Za Zhi. 2002;20(1):1-5.
To explore egg mimotope of Schistosoma japonicum that can be used in the development of diagnostic reagents for schistosomiasis.
By performing three rounds of biopanning in the affinity selection before picking out single clones for identification, target specific phages were effectively enriched from a random 15-peptides phage library with an immobilized mAb 6B12 which is specific to egg antigen of S. japonicum as a bait. By using ELISA, competitive ELISA with natural egg antigen as competitor, and Western blotting, 13 phage clones with high affinity and specificity to 6B12 were obtained and amplified from 400 single clones.
DNA sequencing revealed that all the 13 selected single clones were identical in displaying gene sequence. Serum samples were tested by ELISA for the presence of IgG antibodies to the phages with mimic epitope, and showed that there was a significant difference between healthy volunteers and schistosomiasis patients.
It is possible that the phage display peptide antigen, egg mimotope of S. japonicum may be a replacement of natural egg antigen for the diagnosis of schistosomiasis.
探索日本血吸虫虫卵模拟表位，用于血吸虫病诊断试剂的研发。
以日本血吸虫虫卵抗原特异性单克隆抗体6B12为诱饵，从随机15肽噬菌体文库中通过三轮亲和筛选淘选单克隆进行鉴定，有效富集靶特异性噬菌体。通过酶联免疫吸附测定（ELISA）、以天然虫卵抗原为竞争者的竞争性ELISA以及蛋白质印迹法，从400个单克隆中获得并扩增出13个对6B12具有高亲和力和特异性的噬菌体克隆。
DNA测序显示，所有13个筛选出的单克隆展示的基因序列相同。通过ELISA检测血清样本中针对具有模拟表位噬菌体的IgG抗体的存在情况，结果显示健康志愿者和血吸虫病患者之间存在显著差异。
日本血吸虫噬菌体展示肽抗原——虫卵模拟表位有可能替代天然虫卵抗原用于血吸虫病的诊断。
