华支睾吸虫模拟抗原表位的筛选和鉴定

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。

Screening and identification of mimic antigen epitopes of Clonorchis sinensis.

Objective To screen antigenic epitope of Chonorchis sinensis from phage 12-mer peptide library. Methods Three rounds of affinity selection with immunoglobulin G from 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 immunology 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 discerned by sera from patients infected with Clonorchis sinensis. Conclusion The mimotope of Clonorchis sinensis can be obtained by phage peptide library screening.