摘要
目的 比较两种不同方法筛选噬菌体肽库所获得的日本血吸虫抗原模拟表位的抗原性。方法 以日本血吸虫病患者血清免疫球蛋白 (Sj- Ig)作为靶分子 ,分别以噬菌体 12肽库 (Sj A)和经过蚯蚓免疫兔血清免疫球蛋白 (L - Ig)筛选一轮后的噬菌体 12肽库 (Sj B)为原肽库 ,进行 3轮吸附 -洗脱 -扩增免疫筛选。随机挑取 Sj A和 Sj B蓝色噬菌斑各 2 4个 ,扩增后用 EL ISA方法检测其抗原性。结果 2 4个 Sj A克隆中有 6个可以与 Sj- Ig特异结合 ,其中 A4 91 nm值较高的 2个克隆具有较好的特异性和敏感性 ;2 4个 Sj B克隆中有 10个可以与 Sj- Ig特异结合 ,其中 A4 91 nm值较高的 5个克隆具有较好的抗原性。比较 Sj A和 Sj B两组间克隆的抗原性 ,结果显示 Sj B的目的克隆具有更好的特异性和敏感性。结论 从 Sj A和 Sj B中均筛选到了对日本血吸虫病具有较高潜在诊断价值的抗原模拟表位 ,提示以异源性抗体结合后的肽库作为原肽库来筛选寄生虫抗原模拟表位是可行的 。
Objective To compare the antigenicity of the mimic epitopes of diagnostic antigen of schistosomiasis from phage 12-mer peptide library and phage 12-mer peptide library screened by anti-lumbricus immunoglobulin (L-Ig ). Methods SjA was the original phage 12-mer peptide library and SjB was the phage peptide library immunoscreened by specific L-Ig, which was purified from sera of rabbits. Specific Sj-Ig,purified from sera of patients with schistosomiasis, was used to immunoscreen SjA and SjB, respectively. After three rounds of panning, 24 positive plaques were selected and amplified from SjA and SjB, respectively. ELISA was used to examine the antigenicity of each clone. The sensitivity and the specificity of positive phage clones were confirmed by detecting the sera of different parasitic patients. Results Six phage clones of SjA could bind to the sera of schistosomiasis patients, while ten clones of SjB had a same sensitivity in ELISA. Two of six SjA positive clones showed high immuno-activity while five of six SjB positive clones did. ELISA results showed positive clones of SjB had higher specificity and sensitivity. Conclusion The mimic epitopes obtained both from SjA and SjB played a good role in serodiagnosis of schistosomiasis, which built a good foundation for the further study for obtaining a new diagnostic reagent.
出处
《中国血吸虫病防治杂志》
CAS
CSCD
2004年第3期181-184,共4页
Chinese Journal of Schistosomiasis Control
基金
湖北省重点课题 ( No.WJ0 15 0 5)
关键词
日本血吸虫
蚯蚓
模拟表位
抗原性
噬菌体肽库
Schistosoma japonicum
Lumbricus
Mimic epitope
Antigenicity
Phage peptiden library
