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微生物与感染
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上海2009甲型H1N1流行性感冒病毒全基因组分析
宋志刚1; 田棣1; 张小楠1; 任广旭1,2; 周志统1; 何静1; 袁正宏1,2; 胡芸文1
1. 上海市(复旦大学附属)公共卫生临床中心, 上海201508; 2. 复旦大学上海医学院医学分子病毒学重点实验室, 上海200032
Whole-genome analysis of 2009influenza A virus subtype H1N1 isolates in Shanghai
SONG Zhi-Gang1; TIAN Di1; ZHANG Xiao-Nan1; REN Guang-Xu1,2; ZHOU Zhi-Tong1; HE Jing1; YUAN Zheng-Hong1,2; HU Yun-Wen1
1. Shanghai Public Health Clinical Center Affiliated to Fudan University,Shanghai 200032, China; 2. Key Laboratory of Medical Molecular Virology, Shanghai Medical College, Fudan University, Shanghai 200032, China
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摘要: 2009 年全球暴发2009 甲型H1N1 流行性感冒( 简称流感) 疫情, 上海于2009 年5 月出现第1 例输入型病例。为了解上海地区输入型2009 甲型H1N1 流感病毒的生物学特征, 以上海较早发现的2 例输入型甲型H1N1 流感患者作为研究对象, 分离出A/ Shanghai/ 37T/ 2009 和A/ Shanghai/ 71T/ 2009 病毒, 利用实时定量荧光反转录- 聚合酶链反应( RT- PCR) 鉴定病毒, 通过扫描透射电子显微镜观察、免疫荧光检测、全基因组测序和生物信息软件分析, 对这2 株流感病毒形态、结构、耐药性、基因特点和病毒型别等进行研究。结果显示, 病毒呈现正黏病毒颗粒形态特征; 犬肾( MDCK) 细胞内的病毒能与患者恢复期血清反应。此2 株病毒的全基因核酸序列和氨基酸序列与美国参考株A/ Calif ornia/ 04/ 2009( H1N1) 有较高同源性, 其中第31 位氨基酸残基发生改变。对金刚烷胺耐药, 而对奥司他韦敏感。基于全基因组的系统发育分析, 确认此2 株病毒属2009 甲型H1N1 流感病毒。

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关键词 2009甲型H1N1流行性感冒病毒病毒分离株核酸测序进化树分析    
Abstract:A new strain of influenza A virus subtype H1N1 first identified in April 2009 has caused global pandemics . The first confirmed case in Shanghai was in May 2009. In order to study the characteristics of the 2009 influenza A virus subtype H1N1 in Shanghai , the virus stains A/ Shanghai/ 37T/ 2009 and A/ Shanghai/ 71T/ 2009, which were identified by real-ti me reverse transcriptase-polymerase chain reaction ( RT-PCR) , were isolated fromtwo i mported cases . Then , the characteristics of viral structure , drug  esistance , gene andsubtyping of the two viruses were studied by electron microscopy , i mmunofluorescence technique , genome sequencing , and bioinf ormation  oftware analysis . Virus particles from Madin- Darby canine kidney ( MDCK) cell culture showed that it contained numerous enveloped and highly pleomorphic orthomyxovirus  articles with a diameter ranging from60 nmto 80 nmand surrounded by a fringe of welldefined surface projection . The genome nucleotide sequence and amino acid  equence were then compared . The results showed that the two virus strains and the reference strain A/ Calif ornia/ 04/ 2009( H1N1) were highly homologous . Amantadine resistance occurred at residue 31 ( Asn) in M2 protein, while in the neuraminidase protein domain, it was sensitive to oseltamivir . Phylogenetic analysis showedthat thetwo strains were highly genetically si milar to those previously reported on the 2009 influenza A virus subtype H1N1 strains , suggesting the same origins f or the two Shanghai strains and 2009influenza A virus subtype H1N1 strains .

 

Key wordsInfluenza A virus subtype H1N1    Virus isolate    Nucleotide sequencing    Phylogenetic analysis
通讯作者: 胡芸文; 袁正宏     E-mail: ywhu@shaphc.org; zhyuan@shaphc.org
引用本文:   
宋志刚1; 田棣1; 张小楠1; 任广旭1,2; 周志统1; 何静1; 袁正宏1,2; 胡芸文1. 上海2009甲型H1N1流行性感冒病毒全基因组分析[J]. 微生物与感染 , 2010, 5(4): 225-231.
SONG Zhi-Gang1; TIAN Di1; ZHANG Xiao-Nan1; REN Guang-Xu1,2; ZHOU Zhi-Tong1; HE Jing1; YUAN Zheng-Hong1,2; HU Yun-Wen1. Whole-genome analysis of 2009influenza A virus subtype H1N1 isolates in Shanghai. Journal of Microbes and Infections, 2010, 5(4): 225-231.
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