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目的了解濮阳地区手足口病患者标本中分离的肠道病毒71型(EV71)病毒全基因组特征。方法选取本市发生的7例手足口病患者肠道病毒EV71型核酸阳性标本,使用人横纹肌肉瘤细胞(rhabdomyosarcom cell,RD)进行病毒培养,RT-PCR型别鉴定,对分离的7株EV71型毒株通过12对引物进行分段扩增,扩增产物进行EV71型全基因组序列测定和基因进化特性分析,利用生物信息学软件对基因组序列进行分析并构建EV71病毒分类和发育图谱。结果共分离出7株EV71病毒,系统发育树显示7株病毒在分类上均属于C4亚型C4a分支。在发育相似度上主要分为两大类,其中PY464与宁波2010年NINGBO.CHN/065/2010EV71分离株相似度最高;其他6株呈现聚类,与山东临沂EV71SDLY107分离株相似度最高。在同一时期、同一地区,轻、重症病例分离株VP1区段无明显差别,而全基因区段存在一定差别。结论在濮阳市同一地区、同一时期,亦呈现出以某一种手足口病病毒为主的流行趋势。
Objective To understand the genome-wide characteristics of enterovirus 71 (EV71) virus isolated from specimens of hand-foot-mouth disease in Puyang area. Methods The positive samples of enterovirus EV71 in seven cases of hand, foot and mouth disease in our city were selected. The virus was cultured in rhabdomyosarcom cells (RD) and identified by RT-PCR. Seven strains of EV71 Strain was amplified by 12 pairs of primers. The amplified product was sequenced and analyzed for EV71 genomic sequence analysis. Bioinformatics software was used to analyze the genomic sequence and construct the EV71 virus classification and development map. Results Seven EV71 viruses were isolated and the phylogenetic tree showed that the seven viruses belonged to C4 subtype C4a in the classification. There are two main categories of developmental similarity, PY464 has the highest similarity with Ningbo 2010 NINGBO.CHN / 065 / 2010EV71 isolates; the other six clusters show the highest similarity with Shandong EV71SDLY107 isolates. In the same period, the same region, mild and severe cases of isolates VP1 segment no significant difference, but there are some differences between the whole gene segments. Conclusions In the same area of Puyang City, the same period, also showed a trend of HFMD.