鸭源H5N1亚型禽流感疫苗候选株的构建  

Construction of duck origin H5N1 subtype avian influenza vaccine candidate

作　　者：唐应华[1] 吴培培[1] 彭大新[1] 张文俊[1] 李彦芳[1] 汪文斌[1] 刘秀梵[1] 

机构地区：[1]扬州大学兽医学院农业部畜禽传染病学重点开放实验室,江苏扬州225009

出　　处：《中国兽医科学》2009年第1期54-57,共4页Chinese Veterinary Science

基　　金：国家高技术研究发展计划(863)项目(2006BAA10A205);江苏省高技术研究计划项目(BG2007336)

摘　　要：H5N1亚型clade 2.3.4为我国南方地区的优势毒株。A/mallard/Huadong/S/2005(S)病毒在遗传进化上属于clade 2.3.4,而A/mallard/Huadong/Y/2003(Y)病毒属于clade 2。通过删除S病毒和Y病毒的HA多碱性氨基酸序列,S病毒由PLRERRRKRGL改变为PLREGRGL,Y病毒由PQR-ERRKKRGL改变为PQREGRGL,使之成为低致病性特征的HA基因。将S病毒和Y病毒的NA基因和突变的HA基因分别组合,由A/Puerto Rico/8/34(PR8)病毒提供6个内部基因,进行拯救,共获得4个致弱病毒(SS,SY,YS,YY)。这4个致弱病毒对SPF鸡的静脉内致病指数(IVPI)为0,对10日龄SPF鸡胚无致病性。SS、SY、YS、YY病毒第10代鸡胚尿囊液的HA效价分别为9、11、111、1 lb。Currently,H5N1 viruses in clade 2.3.4 are predominant viruses which prevail in Southern China. Two isolates of H5N1 viruses A/mallard/Huadong/S/2005（S） placing at clade 2.3.4 and A/mal- lard/Huadong/Y/2003（Y） clustering to clade 2 were closely related to the prevalent viruses in China. By deleting the multiple basic amino acid residues at HA cleavage site from S virus and Y virus,the sequence PLRERRRKRGL of S virus and the sequence PQRERRKKRGL of Y virus were changed into PLREGRGL and PQRERRKKRGL,respectively. Four attenuated viruses SS, SY, YY and YS were rescued by reverse genetics,in which NA and mutational HA genes were derived from S or Y viruses,and internal gene was afforded by PR8. The four attenuated viruses were nonpathogenic to SPF chickens（IVPI=0） and 10-dayold embryonated SPF chicken eggs. HA titer of the tenth passage allantoic fluids for SS virus was 9 lb and HA titers for the other three viruses were 11 lb.

关 键 词：H5N1 HA裂解位点 反向遗传 疫苗候选株 

分 类 号：S852.659.5[农业科学—基础兽医学]