甲型流感病毒亚型抗原肽合成及其免疫反应性比较鉴定

    郭 雪1，2 王 琳1，2 杨鹏飞1 咸庆杰1，3 张丽萍1 平芮巾1 胡孔新1

    （1.中国检验检疫科学研究院，北京 100025；2.长春生物制品研究所，长春130000；

    3.潍坊医学院，潍坊 261000）

    摘要 〔目的〕针对甲型流感病毒血凝素（Hemagglutinin，HA）蛋白同一区段不同亚型的合成肽，比较分析其免疫反应性和交叉反应性，获取具有免疫诊断学意义的抗原肽。〔方法〕通过生物信息学分析，针对新甲型H1N1、季节性H1N1、H2N2、H3N2和H5N1共5种亚型甲型流感病毒，选择2个HA肽段区位设计2组抗原肽段（19肽和23肽）合成，每组5条共10条，并分别与钥孔戚血蓝蛋白(KLH)和牛血清白蛋白(BSA)载体偶联形成全抗原后，免疫家兔制备兔源抗体，建立免疫反应性筛选的酶联免疫吸附法(ELISA)，比较合成肽的免疫反应性。〔结果〕以阳性血清平均OD 490  nm /阴性血清平均OD 490  nm （Ｐ/Ｎ）值为判断标准，筛选出具有免疫反应性的2条特异性抗原肽（KYVKSTKLRLAT?鄄GLRNVPS，VKNGTYDYPQYSEEARLNREEIS），前者具有亚型特异反应性，后者则具有一定的亚型通用免疫反应性。〔结论〕筛选出具有免疫反应性合成肽抗原，为进一步建立流感病毒亚型的快速鉴定诊断方法奠定了基础。

    关键词 甲型流感病毒；合成肽；血凝素；免疫反应性；交叉反应性；比较；鉴定

    〔中图分类号〕　R511.7　〔文献标识码〕　B

    Identification and Comparison of theImmunoreactivity for Synthetic Peptide of Influenza A VirusSubtypes   Guo Xue1，2, Wang Lin1，2, Yang Pengfei1, XianQingjie1，3, Zhang Liping1, Ping Ruijin1, Hu Kongxin1. (1.ChineseAcademy of Inspection and Quarantine,Beijing100025,China;2.Changchun Institute of Biological Products,Changchun130000,China;3.Weifang Medical University,Weifang 261000,China.)

    [Abstract]   Objective  To aimat synthetic peptide in different subtypes of the same section ofinfluenza A virus HA protein, and to compare immunoreactivity andcross-reactivity in order to obtain immune diagnosticpeptides.  Method   Basing on bioinformatics takenfrom new influenza H1N1, seasonal H1N1, H2N2, H3N2 and H5N1influenza A virus,five subtypes of influenza A virus, two groups ofsynthesized antigen peptides (19 peptides and 23 peptides) designedby two locations of HA peptide with five pieces were establishedand each was coupled with carriers KLH and BSA to form holoantigenfor immune rabbits antibodies. Then an immune-reactive ELISAscreening test method was set up to detect immune reactivitybetween synthetic peptides.  Result   Using P/Nvalue as criterion, the two specific antigen peptides(KYVKSTKLRLATGLRNVPS, VKNGTYDYPQYSEEARLNREEIS) with immuneresponses were chosen. The former was with subtype-specificreactivity and the latter has a certain type of immune response ingeneral. Conclusion  Immune reactive synthetic peptide antigenwill provide a prerequisite for further rapid diagnosis ofinfluenza subtypes.

    [Key words]   Influenza Avirus；Synthetic peptide；HemagglutininImmunoreactivity；Cross-reactivity; Camparison; Identification