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[2009年北京甲型H1N1流感轻症与重症病例病毒全基因组分析]

[Analysis on the whole genome of the influenza H1N1 virus of the mild and severe cases in Beijing in 2009].

作者信息

Shi Wei-xian, Cui Shu-juan, Lu Gui-lan, Huang Fang, Qian Hai-kun, Wang Quan-yi, Deng Ying

机构信息

Institute for Infectious Disease and Endemic Disease Control, Beijing Center for Disease Control and Prevention, Beijing, China.

出版信息

Zhonghua Yu Fang Yi Xue Za Zhi. 2013 May;47(5):420-6.

Abstract

OBJECTIVE

To explore the characteristics of the whole genome of the influenza H1N1 virus of the mild and severe cases in Beijing.

METHODS

A total of 21 samples of throat swabs were collected from surveillance-designated hospitals between June and December in 2009, including 10 severe cases (4 death cases) and 11 mild cases. RNA of the virus were extracted,and the amplified primers of the whole genome were designed.Reverse transcription and PCR were performed to the RNA and then the PCR product was sequenced by software to analyze the evolution of the viral genes and the variation of the amino acids.

RESULTS

Compared with the reference vaccine strain A/California/07/2009 (H1N1), the genetic nucleotide homology in the eight segments of the pandemic H1N1 virus in Beijing in 2009 was higher than 99%, without significant variation. Among them,the genetic distance of hemagglutinin (HA), neuraminidase (NA) and nucleoprotein (NP) was comparatively far, separately 0.0050, 0.0040 and 0.0040.The gene of HA, P83S, the gene of NA, N248D, the gene of polymerase (PA), P224S and the gene of NP, V100I and L122Q were found to mutate in all the samples. Genes of HA, NA, NP, PA, PB 2 and nonstructural protein (NS1) in severe cases showed obviously clustered evolution. The mutation of gene S128P and S203T of HA, gene R269R and D547E of PA, gene T588I of PB 2 and gene I123V of NS mainly happened in severe cases, separately counting 6, 9, 6, 7, 9 and 6 cases. The relevance between the mutation happened in S203T of HA, R269K and D547E of PA and the severeness of the cases showed statistical significance (P < 0.05). The mutations of HA gene were mainly on the Ca and Cb antigene domains. No drug resistant mutation was found on NA gene but happened on matrix protein 2 (M2 gene). None of the mutations were found on the virulence related genes.

CONCLUSION

A high homology was found between the pandemic H1N1 virus in Beijing in 2009 and the reference vaccine strain A/California/07/2009(H1N1). Mutational sites related with the severe and fatal cases were found, but not the virulence related mutation.

摘要

目的

探讨北京甲型H1N1流感轻症与重症病例的全基因组特征。

方法

于2009年6月至12月期间,从监测定点医院共采集21份咽拭子样本,其中包括10例重症病例(4例死亡)和11例轻症病例。提取病毒RNA,设计全基因组扩增引物。对RNA进行逆转录和PCR,然后通过软件对PCR产物进行测序,分析病毒基因的进化及氨基酸变异情况。

结果

与参考疫苗株A/California/07/2009(H1N1)相比,2009年北京甲型H1N1流感病毒8个基因片段的核苷酸同源性均高于99%,无明显变异。其中,血凝素(HA)、神经氨酸酶(NA)和核蛋白(NP)的遗传距离相对较远,分别为0.0050、0.0040和0.0040。在所有样本中均发现HA基因的P83S、NA基因的N248D、聚合酶(PA)基因的P224S、NP基因的V100I和L122Q发生突变。重症病例中HA、NA、NP、PA、PB2和非结构蛋白(NS1)基因呈现明显的聚类进化。HA基因的S128P和S203T、PA基因的R269R和D547E、PB2基因的T588I和NS基因的I123V突变主要发生在重症病例中,分别为6例、9例、6例、7例、9例和6例。HA基因的S203T、PA基因的R269K和D547E突变与病情严重程度的相关性具有统计学意义(P<0.05)。HA基因的突变主要发生在Ca和Cb抗原结构域。NA基因未发现耐药性突变,但基质蛋白2(M2基因)发生了突变。在毒力相关基因上均未发现突变。

结论

2009年北京甲型H1N1流感病毒与参考疫苗株A/California/07/2009(H1N1)具有较高同源性。发现了与重症和致死病例相关的突变位点,但未发现毒力相关突变。

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