[Molecular Detection and Genomic Characterization of Enterovirus D68 among Children with Severe Acute Respiratory Infection in Beijing and Shanghai].

Yanqun Wang, Yanjie Zhao, Jun Shen, Zhengde Xie, Yamin Li, Gaoshan Liu, Yongliang Lou, Roujian Lu, Wenjie Tan
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Abstract

To understand the prevalence and molecular typing of enterovirus D68 among children with severe acute respiratory infection(SARI)in Beijing and Shanghai,259 respiratory samples were collected from in Beijing during 2008-2010,and 441 respiratory samples were collected in Shanghai city between 2013and2014.All the samples were used for the screening of EV-D68 by nest RT-PCR and sequencing, and then EV-D68-positive samples were used for the complete genome sequencing through overlapping PCR. All available EV-D68full-length genomes collected from GenBank were used for phylogenetic analysis and comparison of EV-D68 types prevalent in China and America. One(0.4%)from 259 respiratory samples in Beijing was positive for EV-D68,and 4(0.9%)among the 441 samples from Shanghai were positive for EV-D68.Phylogenetic analysis of full length genome indicated that the EV-D68 prevalent in Beijing belong to Clade A2 and Clade B2,different from the American popular strains(Clade A1,Clade B1,Clade B4 and Clade B5).Partial sequence analysis declared phylogenetic conflict among different gene sequences. We concluded that the prevalence rate of EV-D68 among SARI Children in Beijing and Shanghai currently was lower(5/700;<1%),and the EV-D68 genotype prevalent in China and America belong to different clusters. Partial sequence analysis indicated that intratypic recombinant events may occur in EV-D68 prevalent in China.

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[北京和上海地区严重急性呼吸道感染儿童肠道病毒D68的分子检测和基因组特征]。
为了解北京和上海地区严重急性呼吸道感染(SARI)患儿肠道病毒D68的流行情况及分子分型,本研究于2008-2010年在北京采集了259份呼吸道样本,2013 - 2014年在上海采集了441份呼吸道样本。所有样本通过巢式RT-PCR筛选EV-D68并测序,然后将EV-D68阳性样本通过重叠PCR进行全基因组测序。利用从GenBank中收集到的所有EV-D68全基因组对中国和美国流行的EV-D68型进行系统发育分析和比较。北京地区259份呼吸道样本中1份(0.4%)EV-D68阳性,上海地区441份(0.9%)EV-D68阳性。全基因组系统发育分析表明,北京流行的EV-D68与美国流行的EV-D68不同,属于A2和B2支系。部分序列分析表明不同基因序列存在系统发育冲突。我们得出结论,目前北京和上海严重急性呼吸道感染儿童中EV-D68的患病率较低(5/700;
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