高增殖性能犬流感CIV-PR8重组病毒的制备  

Development of high-replicated canine influenza virus-PR8 reassortant virus

作　　者：周洁文[1,2] 滕巧泱[2] 闫丽萍[2] 边延峰 张旭[2] 徐大伟[2] 石迎[2] 周雨霞[1] 李泽君[2] 

机构地区：[1]内蒙古农业大学兽医学院,内蒙古呼和浩特010018 [2]中国农业科学院上海兽医研究所,上海200241 [3]天津生机集团股份有限公司,天津300384

出　　处：《中国预防兽医学报》2012年第7期518-522,共5页Chinese Journal of Preventive Veterinary Medicine

基　　金：公益性行业(农业)专项经费(201003012);国家国际科技合作项目(2010DFB33920);中国农业科学院上海兽医研究所中央级公益性科研院所基本科研业务费专项资金项目(2010JB04)

摘　　要：为获得高效表达犬流感病毒(CIV)HA和NA抗原的疫苗株,本实验通过反向遗传操作技术将A/canine/Zhejiang/01/2010(H3N2)犬流感病毒(简称ZJCIV)的HA和NA基因与A/Puerto Rico/8/34(H1N1)病毒(PR8)的6个内部基因进行重组,拯救并获得了一株重组病毒CIV-PR8。分别用含100 EID50的CIV-PR8和ZJCIV感染SPF鸡胚,含2×103TCID50的CIV-PR8和ZJCIV感染MDCK细胞,不同时间段取病毒样品测定血凝效价和TCID50值。两者比较表明,CIV-PR8分别在接种后36 h和48 h时血凝效价达到峰值,鸡胚尿液中效价可达210,为野生病毒株的16倍;细胞上清血凝价可达28,为野生病毒株的4倍;病毒增殖曲线表明通过鸡胚扩增和细胞扩增,CIV-PR8的滴度均高于野生病毒株ZJCIV。结果表明,拯救的CIV-PR8增殖能力明显高于野生病毒株ZJCIV,该重组病毒有望成为犬流感疫苗研制的候选病毒株。Canine influenza is a highly contagious disease caused by canine influenza virus （C1V）. To enhance the expressions of hemagglutination （HA） and neuraminidase （NA） of C1V, a canine-human reassortant influenza viruses C1V-PR8 was generated by reverse genetic technique, which consisted of the segments encoding HA and NA from H3N2 subtype A/canine/ Zhejiang/01/2010 （ZJCIV） and 6 internal gene segments from A/Puerto Rico/8/34 （H1N1） virus （PR8）. The rescued C1V-PR8 was identified by sequence analysis. Virus titrations showed that the HA titer of CIV-PR8 reached peak value of 2 in allantoic fluid within 36 to 48 hours which was 16 times higher than that of ZJC1V, and the HA titer was 28 in culture medium which was 4 times higher than that of ZJCIV when inoculated in chicken embryos with 100 EIDso and in MDCK cells with 2 x 103 TCIDso of CIV-PR8 and ZJC1V, respectively. The results suggested that reassortant virus of C1V-PR8 was a promising vaccine candidate for canine influenza.

关 键 词：犬流感 重组犬流感病毒 PR8 血凝效价 病毒滴度 

分 类 号：S852.65[农业科学—基础兽医学]