三门峡市手足口病病原分离检测及肠道病毒71型VP1基因分析

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了解三门峡市2011年手足口病例及健康儿童肠道病毒型别及生物学特征。本研究采集55份手足口病临床病例及60份健康儿童粪便接种敏感细胞进行肠道病毒分离,提取RNA,用荧光RT-PCR检测肠道病毒通用型(Pan-enterovirus,PE)、肠道病毒71型(Human enterovirus 71,EV71)、柯萨奇病毒A16型(Coxsackie virus A16,CA16)。EV71、CA16分别用各自的VP1引物全序测定鉴定,对其它肠道病毒使用引物对012/011、008/013进行扩增测序,并与已知序列对比分型鉴定,并且对EV71病毒VP1基因特征进行了分析。临床病例肠道病毒检出率为52.73%(29/55)、健康儿童肠道病毒检出率为18.33%(11/60);临床病例检出22株EV71、4株CA16和3株其它肠道病毒,健康儿童检出肠道病毒11株,其中柯萨奇B组病毒占81.82%(9/11)。对19株EV71病毒VP1基因分析证实,三门峡市2011年流行株为C4a亚群,其进化聚类以灵宝市、卢氏县聚类最为明显。引起手足口病主要为EV71和CA16,健康儿童中存在大量的柯萨奇B组病毒,隐性感染广泛存在,2011年本地EV71病毒流行株为C4a亚群,有明显地域聚集特点。 To understand the type and biological characteristics of enterovirus in hand, foot and mouth disease cases and healthy children in Sanmenxia City in 2011. In this study, 55 cases of hand-foot-mouth disease clinical cases and 60 healthy children stool vaccinated sensitive cells were collected for enterovirus isolation, RNA extraction, detection of Pan-enterovirus (PE), enterovirus 71 (EV71), Coxsackie virus A16 (CA16). EV71 and CA16 were sequenced and identified by their respective VP1 primers. The other enteroviruses were amplified by PCR using the primers 012/011 and 008/013, and were identified by comparison with known sequences. The EV71 and VP1 genes Features were analyzed. The detection rate of enterovirus was 52.73% (29/55) in clinical cases and 18.33% (11/60) in healthy children. In clinical cases, 22 strains of EV71, 4 strains of CA16 and 3 strains of other intestine 11 strains of enterovirus were detected in pathogenic and healthy children, of which 81.82% (9/11) in coxsackie B virus. The analysis of VP1 gene of 19 strains of EV71 virus confirmed that the epidemic strain of C4a in Sanmenxia was C4a subpopulation in 2011, and the clustering was most obvious in Lingbao and Lushi counties. Causing hand-foot-mouth disease mainly EV71 and CA16, there are a large number of Coxsackie B virus in healthy children, latent infection is widespread. In 2011, the local EV71 virus was C4a subpopulation, with obvious regional aggregation.
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