甲型H1N1流感病毒在季流病毒、禽流感病毒共感染时变异可能性的验证

No mutation of influenza A(H1N1) virus at co-infection with seasonal influenza and avian influenza viruses in miniature pigs

目的甲型H1N1流感病毒A/California/7/2009分别与A/Brisbane/10/07和A/ShenZhen/406H/06共感染小型香猪,预测甲流病毒在与季流H3N2病毒/甲流病毒与禽流感病毒共感染时是否会发生变异。方法分别将A/California/7/2009(CA7)与A/Brisbane/10/07(H3N2),A/California/7/2009与A/Shenzhen/406H/06(H5N1)对5～6月龄小型猪共感染,小型猪经复方氯胺酮0.1 mL/kg麻醉后进行滴鼻感染,感染后第5天安乐死动物,取动物肺组织作病毒测序分析。结果 A/California/7/2009(CA7)与A/Brisbane/10/07(H3N2)共感染后,A/California/7/2009病毒PB1基因993位G→A突变,PA基因1659位G→A突变,没有氨基酸的变异。A/California/7/2009与A/Shenzhen/406H/06(H5N1)共感染后A/California/7/2009病毒PB2基因1711位T→C突变。碱基的突变未引起氨基酸的变异。结论 A/California/7/2009(CA7)与A/Brisbane/10/07(H3N2),A/California/7/2009与A/Shenzhen/406H/06(H5N1)共感染后在猪的体内没有发生病毒重组、变异。

Objective To co-infect miniature pigs with influenza A virus A/California/7/2009 with A/Brisbane/10/07 and A/Shenzhen/406H/06 virus,respectively,and to verify whether the co-infection of A/California/7/2009（CA7） and A/Brisbane/10/07（H3N2） or A/California/7/2009 and A/Shenzhen/406H/06（H5N1） influenza viruses will induce mutation in influenza virus.Methods To co-infect the 5-6 month old miniature pigs with A/California/7/2009（CA7） and A/Brisbane/10/07（H3N2） or A/California/7/2009 and A/Shenzhen/406H/06（H5N1） viruses by intranasal instillation after 0.1 mL/kg compound ketamine anesthesia.After 5 days,the animals were euthanized and the lung tissue samples were taken for viral sequencing.Results In the A/California/7/2009（CA7） and A/Brisbane/10/07（H3N2） co-infected pigs,A/California/7 / 2009 virus PB1 993 gene showed G→A and PA Gene 1659 G→A mutations,with no amino acid variation.In the A/California/7/2009 and A/Shenzhen/406H/06（H5N1） co-infected pigs,the A/California/7/2009 virus PB2 gene 1711 showed T→C mutation,with no amino acid variation.Conclusions Our findings demonstrate that no virus recombination and mutation occur after A/California/7/2009（CA7） and A/Brisbane/10/07（H3N2）,A/California/7/2009 and A/Shenzhen/406H/06（H5N1） virus co-infection in the miniature pigs.