| 陈寅,茅海燕,李榛,徐昌平,高见,冯燕,王欣莹,严菊英,张严峻.浙江省人感染H7N9禽流感病毒的基因组序列分析[J].Chinese journal of Epidemiology,2013,34(6):604-608 |
| 浙江省人感染H7N9禽流感病毒的基因组序列分析 |
| Genomic sequences of human infection of avian-origin influenza A (H7N9) virus in Zhejiang province |
| Received:April 20, 2013 Revised:June 26, 2012 |
| DOI： |
| KeyWord: H7N9禽流感病毒 基因组 序列分析 进化树 |
| English Key Word: H7N9 avian influenza virus Genome Sequence analysis Phylogenetic tree |
| FundProject:浙江省自然科学基金;浙江省重点科技创新团队计划;浙江省医学重点学科群建设计划资助 |
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| Abstract: |
| 目的分析浙江省2013年4月初一例人感染甲型H7N9禽流感患者的病原学和基因组序列特征.方法提取患者标本的病毒RNA并用荧光定量RT-PCR方法检测,一步法RT-PCR扩增H7N9禽流感病毒基因组的8个片段,测序并拼接出基因组序列.下载目前已经公布的H7N9禽流感病毒和其他H7、N9亚型的病毒的HA和NA序列,采用Mega 5.1软件对其进行序列比对并构建进化树.根据测序的结果分析该例H7N9禽流感病毒的序列变异情况.结果该患者标本甲型流感、H7亚型和N9亚型均为阳性,通过扩增基因组各片段并进行测序.对血凝素和神经氨酸酶基因进行比对和进化树构建,结果显示该H7N9病毒HA基因与A/duck/Zhejiang/12/2011(H7N3)的亲缘关系最近,NA基因与A/wild bird/Korea/A14/2011 (H7N9)的亲缘关系最近.编码内部蛋白的6个基因均与中国大陆近两年的H9N2毒株最为相似.该标本的病毒HA蛋白发生了Q226L突变,而与已经公布的人感染H7N9禽流感毒株均不一致的是PB2未发生E627K突变.结论从该份临床标本完成了检测和基因组各片段的扩增与测序.该病毒与已报道的人感染H7N9禽流感病毒同源性高,存在Q226L等重要位点的突变,但PB2未发生E627K突变. |
| English Abstract: |
| Objective To analyze the etiology and genomic sequences of human infection of avian-origin influenza A (H7N9) virus from Zhejiang province.Methods Viral RNA was extracted from patients of suspected H7N9 influenza virus infection and real-time RT-PCR was conducted for detection of viral RNA.All 8 segments of influenza virus were amplified by one-step RT-PCR and genomic sequences were assembled using the sequencing data.All the currently available HA and NA genes of the novel H7N9 virus,some other HAs from H7 subtype and NAs from N9 subtype were downloaded from public database for phylogenetic analysis,using the Mega 5.1 software.Mutations and variations were analyzed,using the genomic sequence data.Results Reactions for influenza type A,subtype H7 and subtype N9 were all positive and all the genomic fragments were amplified for sequencing.After alignment,sequences were subjected for phylogenetic analysis.The results revealed highest homology with A/duck/Zhejiang/12/2011 (H7N3) in HA gene and with A/wild bird/Korea/ A14/2011 (H7N9) in NA gene of the H7N9 influenza virus.All 6 genes coding for internal proteins shared highest identities with H9N2 avian influenza which were circulated in the Chinese mainland,in the last two years.The sequenced virus showed Q226L mutation in HA protein,but E627K was not presented in PB2 protein of this virus.The E627K mutation was shared by all the other novel H7N9 viruses resulted in human infections through analysis on the currently available sequences.Conclusion Using the clinical samples,both detection of the viral genes and amplification of all 8 segments of the novel H7N9 influenza virus were accomplished.High homology of the novel H7N9 influenza viruses was observed by phylogenetic test,using the currently available sequences.The virus showed Q226L mutation on HA protein but E627K did not present on PB2 protein of this virus. |
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