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为了解肠病毒D68(EV-D68)在北京、上海地区严重急性呼吸道感染(SARI)儿童中的流行情况和分子特征。本研究采集2008~2010年北京儿童医院SARI儿童呼吸道样品259份,2013~2014年上海复旦大学附属儿科医院SARI儿童呼吸道样品441份,采用套式RT-PCR结合测序方法检测筛选EV-D68阳性样品,设计EV-D68全基因组通用引物,对EV-D68阳性样品进行全基因组扩增,收集NCBI核酸数据库中目前所有的EV-D68全基因组序列进行序列分析,比较EV-D68中国和美国流行株的差异。结果发现:北京地区检测EV-D68阳性样品1例,阳性率为0.4%,上海地区检测EV-D68阳性样品4例,阳性率为0.9%。结合目前中国已上传的所有EV-D68相关序列,分析表明:目前中国地区流行的EV-D68主要属于Clade A2或Clade B2亚型,不同于美国的Clade A1、Clade B1、Clade B4和Clade B5亚型,另外EV-D68部分基因进化分析时出现树形结构冲突现象;上述结果表明目前EVD68在北京、上海地区的SARI儿童群体中的感染率较低(5/700;<1%);中国与美国流行的EV-D68毒株属于不同的分支,差异较大,部分序列分析表明EV-D68病毒可能存在组内重组现象。
To understand the prevalence and molecular characteristics of enterovirus D68 (EV-D68) in children with severe acute respiratory infection (SARI) in Shanghai and Shanghai. This study collected 259 SARI children’s respiratory samples from Beijing Children’s Hospital from 2008 to 2010 and 441 SARI children’s respiratory samples from 2013 to 2014. The positive samples of EV-D68 were screened by nested RT-PCR and sequencing method , Designed EV-D68 genome-wide universal primers, whole genome amplification of EV-D68 positive samples collected all the current EV-D68 genome sequence of NCBI nucleic acid database for sequence analysis, compared EV-D68 China and the United States epidemic strains difference. The results showed that in Beijing, 1 EV-D68 positive sample was detected, the positive rate was 0.4%. In Shanghai, 4 EV-D68 positive samples were detected, the positive rate was 0.9%. Combined with all the EV-D68 related sequences that have been uploaded in China at present, the analysis shows that EV-D68 currently prevailing in China mainly belongs to Clade A2 or Clade B2 subtype, which is different from Clade A1, Clade B1, Clade B4 and Clade B5 in the United States Type, and other structural evolutionary analysis of EV-D68 part of the tree structure of the phenomenon of conflict; the above results show that EVD68 in Beijing, Shanghai SARI children in the lower the prevalence (5/700; <1%); China and The prevalence of EV-D68 strain in the United States belongs to different branches, and the difference is quite large. Partial sequence analysis shows that recombination phenomenon may occur in EV-D68 virus.