期刊文献+

H5与H7亚型流感流行株嵌合病毒株的拯救及初步鉴定

Rescue and preliminary identification of influenza reassortment virus strains of the H5 and H7 subtypes
收藏 分享 导出
摘要 目的探讨H5与H7亚型流感流行株嵌合病毒的拯救并进行鉴定,为新型流感疫苗的研发奠定基础。方法采用脂质体转染的方法,首先依照A型流感病毒A/PR/8/34 HA嵌合片段的设计对H5 2.3.4.4谱系HA和H7亚型的HA进行改造,利用反向遗传技术拯救A/B型嵌合减毒株,对拯救成功的病毒株进行形态学和分子生物学鉴定,并测定病毒滴度;从中选择H5亚型嵌合减毒株以104EID50/50μl的剂量滴鼻感染BALB/c小鼠,观察、记录小鼠的体重变化及死亡情况,并测定攻毒后第3 d及第6 d小鼠肺脏病毒复制能力。结果成功拯救出2种A/B型嵌合减毒株,分别命名为rA/B-H5-NS110和rA/B-H7-NS110,测序显示两株嵌合减毒株的序列与预期一致,并在电镜下观察到了流感病毒样粒子。rA/B-H5-NS110和rA/B-H7-NS110株在MDCK细胞上的病毒滴度均为104.50TCID50/ml;在鸡胚上的病毒滴度rA/B-H5-NS110为105.25EID50/ml,rA/B-H7-NS110株为105.44EID50/ml。小鼠致病性实验中H5亚型嵌合病毒感染组小鼠与对照组相比体重无明显下降,攻毒后小鼠存活率100%;攻毒后第3 d可在小鼠肺脏内检测到嵌合减毒株rA/B-H5-NS110的轻微复制,第6 d时未检测到病毒复制。结论成功地拯救出2株包含有H5 2.3.4.4谱系HA基因以及H7N9流感病毒HA基因的嵌合减毒株,为B型流感病毒包装机制的研究以及新型流感疫苗的研发提供了新的思路。 Objectives To explore the rescue and identification of a chimeric virus of the H5 and H7 subtypes of influenza strains causing outbreaks and to lay the foundation for the development of new influenza vaccines.Methods Liposome transfection was used.First,HA of the 2.3.4.4 clade of the H5 subtype and H7 subtype was specifically altered according to the design of a chimeric fragment of HA of influenza A virus A/PR/8/34.Reverse genetics was used to rescue A/B chimeric attenuated strains.Rescued strains were identified based on morphology and molecular biology,and the viral titer was determined.The chimeric attenuated strain of the H5 subtype was used to infect BALB/c mice via nasal drip at a dose of 104 EID50/50μl.Changes in body weight and the death of mice were observed and recorded,and the lungs were sampled and screened for the ability of the virus to replicate on day 3 and day 6 after challenge.Results Two types of A/B chimeric strains designated rA/B-H5-NS110 and rA/B-H7-NS110 were successfully rescued.Sequencing results indicated that the sequence of the two chimeric attenuated strains was consistent with expectations,and influenza virus-like particles were observed in electron microscopy.The viral titer of the rA/B-H5-NS110 and rA/B-H7-NS110 strains in MDCK cells was 104.50TCID50/ml.The viral titer of the rA/B-H5-NS110 strain in chicken embryos was 105.25 EID50/ml.The viral titer of the rA/B-H7-NS110 strain in chicken embryos was 105.44EID50/ml.The results of pathogenicity testing in mice indicated that there was no significant decrease in body weight of mice in the experimental group infected with the H5 subtype of the chimeric virus,compared to the control group.The survival rate of mice after challenge was 100%.Slight replication of chimeric attenuated strain rA/B-H5-NS110 was detected in the lungs of mice on day 3 after challenge.No titers were detected on day 6.Conclusion Two chimeric attenuated strains with H5 2.3.4.4 lineage HA and H7 N9 HA were successfully recused,providing a new avenue of thought f
作者 赵梦琳 孙伟洋 张醒海 宫语晨 毕津豪 张亚敏 李元果 冯娜 王铁成 王化磊 赵永坤 杨松涛 夏咸柱 高玉伟 ZHAO Meng-lin;SUN Wei-yang;ZHANG Xing-hai;GONG Yu-chen;BI Jin-hao;ZHANG Ya-min;LI Yuan-guo;FENG Na;WANG Tie-cheng;WANG Hua lei;ZHAO Yong-kun;YANG Song-tao;XIA Xian-zhu;GAO Yuwei(Institute of Military VeterinaryMedichie ,Academy of Military Medical Science,Changchun ,China 130122;Key Laboratory of Jilin Province forZoonosis Prevention and Control;Jilin Uriiversity;Jilin Agricultural University;Henan University)
出处 《中国病原生物学杂志》 CSCD 北大核心 2019年第4期381-385,393共6页 Journal of Pathogen Biology
基金 国家重点研发计划项目(No.2016YFDO500203,2016YFD0501004)。
关键词 B型流感病毒 致病性 反向遗传学 Influenza B virus pathogenicity reverse genetics
作者简介 赵梦琳(1993-),女,河北邢台人,硕士,主要从事病毒学与人兽共患病学研究。E-mail:284197767@qq.com;通讯作者:夏咸柱,Email:xiaxzh@cae.cn;通讯作者:高玉伟,Email:yuwei0901@outlook.com.
  • 相关文献

参考文献1

投稿分析

相关作者

内容加载中请稍等...

相关机构

内容加载中请稍等...

相关主题

内容加载中请稍等...

浏览历史

内容加载中请稍等...
;
使用帮助 返回顶部 意见反馈