微生物学报Acta Microbiologica Sinica 48(6)：745~749; 4 June 2008
基金项目: 国家科技支撑计划(2006BAD06A01); 江苏省高校重大基础研究项目(05KJA23016)
*通讯作者。Tel: +86-514-87991416; E-mail: xfliu@yzu.edu cn
作者简介: 唐应华(1981. ), 男, 湖南东安人, 博士研究生, 研究方向为动物性病毒致病分子机理研究。E-mail: tyhlw@yahoo.com.cn
唐应华，吴培培，彭大新，龙进学，张评浒，张文俊，李彦芳，汪文斌，刘秀梵*
(扬州大学兽医学院，农业部畜禽传染病学重点开放实验室, 扬州 225009)
摘要：【目的】为了探讨高致病性禽流感病毒对水禽致病性差异的分子致病机理。【方法】我们对从野鸭分离到的H5N1 亚型禽流感病毒的生物学特性进行鉴定，其中A/mallard/Huadong/Y/2003(Y)是对麻鸭无致病性病毒，而 A/mallard/Huadong/S/2005(S)是对麻鸭高致病性病毒。利用反向遗传技术构建一系列单个和多个基因组合替换基因重排病毒，并验证重排病毒在麻鸭上的致病力。【结果】研究表明，PB2, PB1, PA(3P), HA 单基因以及3P 基因组合替换的使S 病毒对麻鸭的毒力完全致弱，但相应的基因替换后仅使Y 病毒对麻鸭的毒力略有上升。两病毒的其它基因对毒力影响较小。【结论】H5N1 亚型禽流感病毒对麻鸭的致病力受多基因调控，且这种调控作用在不同病毒骨架上的影响不一致，强毒受影响程度远比弱毒的大。
关键词：H5N1；致病力；反向遗传；麻鸭
中图分类号：Q78 文献标识码：A 文章编号：0001-6209 (2008) 06-0745-05
Virulence Changes of H5N1 Avian Influenza Virus after Gene Reassortment
Yinghua Tang, Peipei Wu, Daxin Peng, Jinxue Long, Pinghu Zhang,
Wenjun Zhang, Yanfang Li, Wenbin Wang, Xiufan Liu*
(Key Laboratory of Animal Infectious Disease, Ministry of Agriculture, School of Veterinary Medicine Yangzhou University, Yangzhou 225009, China)
Abstract: [Objective] Recently H5N1 subtype avian influenza virus (AIV) is capable of mortality to aquatic bird. The molecular basis for the virulence of this virus is still poorly understood. [Methods] We characterized two H5N1 subtype viruses, A/mallard/Huadong/Y/2003 (Y) is nonpathogenic to mallard whereas A/mallard/Huadong/S/2005 (S) is highly pathogenic to mallard. Using reverse genetics, we constructed a series of single-gene or multiple-gene reassortants from these two viruses. [Results] Substitution of single-gene for PB2, PB1, PA (3P), HA and of combination for 3P gene resulted in complete attenuation of S virus in mallard. However, these corresponding substitutions only slightly increased virulence of Y virus in mallard. Other gene segments had little contribution to the virulence of both viruses. [Conclusion] These results indicate that the pathogenicity of H5N1 AIV to mallard was regulated by multiple gene segments, and these regulations had more sensitive effect on highly pathogenic virus backbone than on low pathogenic virus backbone.
Keywords: H5N1; pathogenicity; reverse genetics; mallard
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