两种不同噬菌体筛选方法所获的卫氏并殖吸虫抗原模拟表位抗原性比较

目的比较从兔抗蚯蚓血清免疫球蛋白(E-Ig)结合后的噬菌体12肽库中和从噬菌体肽库中筛选出的两种卫氏并殖吸虫抗原模拟表位的抗原性。方法以肺吸虫病患者血清免疫球蛋白(Pw-Ig)作为靶分子,分别对噬菌体12肽库(PwA)和经过 E-Ig 筛选一轮后的肽库(PwB)进行3轮吸附-洗脱-扩增的淘筛,随机挑取 PwA 和 PwB 蓝色噬菌斑各24个扩增,用 ELISA 方法检测并比较两者的抗原性。结果24个 PwA 克隆中有12个可以与肺吸虫病患者血清发生免疫反应,其中 A_(491)nm 值较高的2个克隆具有较好的特异性和敏感性;24个 PwB 克隆中有10个可以被卫氏并殖吸虫病患者血清所识别,其中 A_(491)nm 值较高的2个克隆具有较好的抗原性。结论从 PwA 和 PwB 中均筛选到了对肺吸虫具有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源肽库来筛选寄生虫抗原模拟表位是可行的,为研制新型诊断试剂提供了一条新的着眼点。

Comparing the antigenicity of two kinds of antigen-mimic epitopes of Paragonimus westermani screened from the 12-mer phage peptide library used different methods

Objective To compare the antigenicity of two kinds of mimic epitopes of Paragonimus westermani,which were sreened from phage 12-mer peptide library and phage 12-mer peptide library screened by anti-lumbricus immunoglobulin(E-Ig)of rabbits respectively.Methods The original phage pep- tide library was named PwA.The phage peptide library,which was immunoscreened by specific E-Ig purified from sera of rabbits,was called PwB.And specific Pw-Ig,purified from sera of Paragonimiasis patients,was used to immunoscreen PwA and PwB in three rounds.After 3 rounds of panning,24 positive plaques from PwA and PwB were selected and amplified,respectively.ELISA was used to examine the immunoactivity of each clone.Finally the antigenicity of positive clones was analysed between PwA and PwB.Results 12 phages clones of PwA could bind to the sera of Paragonimiasis patients,while 10 clones of PwB had a same sensitivity in ELISA.Both 2 of 6 PwA and PwB positive colones,respectively,showed high immuno-activity. Conclusion These two kinds of mimic epitopes obtained played a good role in serodiagnosis of Paragonimus westermani.The results showed that it was available to get diagnostic mimic epitopes of Paragonimiasis,im- munoscreened from peptide library screened by E-Ig.