3株H9N2亚型重组禽流感病毒生物学特性及免疫原性研究  

Biological Characteristics and Immunogenicity of Three H9N2 Subtype Recombinant Avian Influenza Viruses

作　　者：栾璐 倪小舒 刁小龙 刘云迎 印广浩 何海蓉 LUAN Lu;NI Xiaoshu;DIAO Xiaolong;LIU Yunying;YIN Guanghao;HE Hairong(l.Key Laboratory of Safety Assessment of Agricultural Products,Livestock and Poultry Inputs,Ministry of Agriculture and Rural Affairs,Zhongchong Sino Biotech Taizhou Co.,Ltd.,Taizhou,Jiangsu 225300;Wuxi Zhengzheng Precision Medical Laboratory Co.,Ltd.,Wuxi,Jiangsu 214000)

机构地区：[1]中崇信诺生物科技泰州有限公司,农业农村部农产品畜禽投入品安全评价重点实验室,江苏泰州225300 [2]无锡正则精准医学检验有限公司,江苏无锡214000

出　　处：《中国家禽》2022年第3期38-43,共6页China Poultry

基　　金：“十三五”国家重点研发计划项目(2017YFD0500701-6)。

摘　　要：为评估经反向遗传技术构建的3株H9N2亚型重组禽流感(AIV)病毒A3-PR8株、AH515-PR8株和TM343-PR8株的生物学特性和免疫原性,研究将3株重组毒株在鸡胚上连续传代,3株重组毒株均能在鸡胚中稳定增殖,红细胞凝集价达到9.0 log2。与对应的亲本分离株相比,重组毒株的病毒含量(EID_(50))提升0.5~0.8个滴度。以3株重组毒株为抗原制备油乳剂灭活疫苗,0.2 mL/只接种21日龄SPF鸡,免疫后第7、14、21、28天采血,测定血清中H9亚型AIV的血凝抑制试验(HI)抗体效价,结果显示,免疫后28 d三种疫苗免疫鸡几何平均HI效价都可达8.0 log_(2),AH515-PR8株免疫组抗体效价最高达8.8 log_(2)。免疫后28 d分别以10^(6.0) EID_(50)的剂量进行H9N2 AIV WJ57株的攻毒保护试验,在攻毒后,AH515-PR8和TM343-PR8组免疫鸡均不发病、不死亡,排毒量有明显下降,其中AH515-PR8株免疫组排毒量最低,5 d病毒分离率为0。研究表明,经反向遗传技术构建的AH515-PR8株相较于TM343-PR8株和A3-PR8株,更具有生长繁殖能力和强免疫原性的特点,为H9N2亚型禽流感疫苗的应用和开发提供了参考。To estimate the biological characteristics and immunogenicity of three recombinant avian influenza H9N2 subtype(AIV) virus strains A3-PR8,AH515-PR8 and TM343-PR8 constructed by reverse genetic technique,the three recombinant strains were passed continuously and proliferated stably in chicken embryos.Besides,the erythrocytes agglutination value reached 9 log;.The content(EID_(50)) of the recombinant strain increased by 0.5 to 0.8 titers compared with the corresponding parent isolates.The oil emulsion inactivated vaccine was prepared with three recombinant viruses as antigen.The SPF chickens were inoculated with 0.2 mL/chicken at 21 days of age.Blood samples were collected on 7 th,14 th,21 st and 28 th days after immunization,to determine the antibody titer(HI) of H9 subtype in serum.The results showed that the geometric average HI titer of the three vaccines all reached 8.0 log;on the 28 th day after immunization,and the highest value of AH515-PR8 was 8.8 log;.At 28 th day after immunization,the challenge-protection test of H9N2 AIV subtype(strain WJ57) was conducted with 106.0 EID_(50) respectively.The results indicated that the chicken immunized with AH515-PR8 and TM343-PR8 had no symptoms of morbidity or death,and the amount of virus excretion decreased significantly.Especially,strain AH515-PR8 had the lowest detoxification capacity,and the virus isolation rate was 0 in 5 th day.The conclusion could be drawn that using Texas/05/2009×Puerto Rico/8/1934 strain of H1 N1 subtype influenza virus as the internal gene donor,the strain AH515-PR8 constructed by reverse genetic technology had the characteristics of reproductive capacity and stronger immunogenicity compared with TM343-PR8 and A3-PR8,which provided a reference for the application and development of H9N2 subtype avian influenza.

关 键 词：重组禽流感病毒 油乳剂灭活疫苗 免疫效力试验 

分 类 号：S852.659.5[农业科学—基础兽医学]