Yan Ju-Ying, Chen Yin, Li Zhen, Gong Li-Ming, Lu Yi-Yu, Zhang Yan-Jun
Zhejiang Provincial Center for Disease Control and Prevention, Hangzhou 310051, China.
Bing Du Xue Bao. 2011 Sep;27(5):421-6.
To identify and trace the pathogen of acute hemorrhagic conjunctivitis (AHC) epidemic in Zhejiang Province in 2010. Viral nucleic acid of Enterovirus (EV) and Coxsackievirus A24 variant (CA24v) were directly detected by real-time RT-PCR from the conjunctival swab collected from suspected patients. The virus was isolated from the swab samples using Hep-2 cell. The viral RNAs were extracted from the isolated viruses and followed by RT-PCR to amplify VP1 gene and 3C protease region(3C). The amplified fragments were sequenced and phylogenetic trees were also constructed. Eight out of 13 swab samples from suspected patients were both positive for EV and CA24v RNA (61.5%), 6 CA24v strains were isolated (46.2%). The complete VP1 genes of CA24v in 4 sequenced virus strains were 915 nt in length and the complete 3C genes were 549 nt in length. All VP1 and 3C genes were confirmed without any insertion or deletion. The identity of nucleotide and amino acid in 3C between the 2010 isolated strains and the prototype strain EH24/70 were 85.2%-85.8% and 96.2%-96.7%, and that between the 2010 Zhejiang strains and the Zhejiang,Yunnan and Guangdong CA24v strains isolated between 2007-2008 were 93.4%-93.8% and 96.7%-97.3%, respectively. The phylogenetic tree of 3C indicated that the isolated CA24v viruses of Zhejiang in 2010 located in the CA24v IV genotype cluster 4 (GIV-C4) and all the VP1 genes located in the human Enterovirus C (EV-C) CA24v. These findings indicated that AHC epidemic in Zhejiang Province in 2010 was caused by CA24v GIV-C4 viruses and they most likely evolved from CA24v viruses circulating locally in external environment from 2002.
鉴定并追踪2010年浙江省急性出血性结膜炎（AHC）疫情的病原体。采用实时逆转录聚合酶链反应（real-time RT-PCR）直接从疑似患者结膜拭子中检测肠道病毒（EV）和柯萨奇病毒A24变异株（CA24v）的病毒核酸。使用Hep-2细胞从拭子样本中分离病毒。从分离出的病毒中提取病毒RNA，随后进行逆转录聚合酶链反应扩增VP1基因和3C蛋白酶区（3C）。对扩增片段进行测序并构建系统发育树。13份疑似患者拭子样本中有8份同时检测到EV和CA24v RNA呈阳性（61.5%），分离出6株CA24v毒株（46.2%）。4株测序病毒株中CA24v的完整VP1基因长度为915个核苷酸，完整3C基因长度为549个核苷酸。所有VP1和3C基因均确认无任何插入或缺失。2010年分离株与原型株EH24/70的3C基因核苷酸和氨基酸同一性分别为85.2% - 85.8%和96.2% - 96.7%，2010年浙江毒株与2007 - 2008年分离的浙江、云南和广东CA24v毒株的同一性分别为93.4% - 93.8%和96.7% - 97.3%。3C基因的系统发育树表明，2010年浙江分离的CA24v病毒位于CA24v IV基因型簇4（GIV-C4），所有VP1基因均位于人肠道病毒C（EV-C）CA24v。这些发现表明，2010年浙江省AHC疫情由CA24v GIV-C4病毒引起，它们很可能由2002年在外部环境中本地传播的CA24v病毒进化而来。
