鸭源H5亚型禽流感全禽源分子标记疫苗候选株的构建  

Generation of Duck Origin H5 Subtype Avian Influenza Vaccine Candidate with Heterogenous NA Gene

作　　者：张文俊[1] 薛涛[1] 吴小伟[1] 唐应华[2] 彭大新[1] 刘秀梵[1] 

机构地区：[1]扬州大学农业部畜禽传染病学重点开放实验室,扬州225009 [2]江苏省农业科学院国家兽用生物制品工程技术研究中心,南京210014

出　　处：《畜牧兽医学报》2011年第5期685-691,共7页ACTA VETERINARIA ET ZOOTECHNICA SINICA

基　　金：现代农业产业技术体系建设专项资金(nycytx-41-G07);国家重点基础研究发展计划(973计划)(2011CB505003)

摘　　要：为了开发用于南方水禽且适合血清学监测的H5亚型禽流感疫苗,作者通过反向遗传技术,删除A/mal-lard/Huadong/S/2005(H5N1,S株)病毒HA编码多碱性氨基酸序列,使之成为低致病特征,分别与A/duck/Eng-land/1/1956(H11N6,E株)和A/Chicken/Shanghai/F/98(H9N2,F株)的NA组合,再以本室禽源高产病毒F株的6个内部片段为骨架,构建全部基因都来自禽流感的疫苗株。成功拯救出2株重组病毒,分别命名为rH5N6/F和rH5N2/F,并引入分子标记N6和N2。重组病毒在鸡胚和MDCK细胞上均具有较好的繁殖能力,且rH5N6/F更适合在鸡胚中生产,对SPF鸡和鸡胚无致病性。重组病毒在clade2.3.4毒株中具有很好的抗原代表性,引入的分子标记有利于血清学监测的区分,为防控水禽H5亚型禽流感提供了良好的疫苗候选株。The aim of this study was to develop H5 subtype avian influenza vaccine with all genes from avian origin influenza viruses,and provide a DIVA（Differentiating Infected from Vaccinated Animals） strategy for the control of avian influenza.Sequences encode several basic amino acids of HA gene from A/mallard/Huadong/S/2005（H5N1,S strain） were deleted.Using reverse genetics system,we rescued two reassortant viruses（rH5N6/F and rH5N2/F） that contained NA genes from A/duck/England/1/1956（H11N6,E strain） or A/Chicken/Shanghai/F/98（H9N2,F strain） and internal genes from F strain.The two vaccine viruses were avirulent to SPF chickens and embryonated chicken eggs,replicating well in eggs and MDCK cells,while rH5N6/F showed better growth in eggs.The results indicate that the reassortant viruses are suitable for vaccine manufacturing and being used as reference vaccine virus against the H5N1 clade 2.3.4 viruses in China.

关 键 词：禽流感病毒 反向遗传 H5 DIVA疫苗 

分 类 号：S852.659[农业科学—基础兽医学]