摘要
目的分析引起神经症状的柯萨奇病毒A组16型病毒（coxsackievirus A16, CA16）VP1基因特征。方法采集表现神经症状的病例临床标本及信息，采用荧光RT-PCR方法检测病原，肠道病毒阳性标本进行病毒分离培养，培养阳性标本采用RT-PCR方法扩增VP1序列并测序。利用DNAStar 5.0和Mega 5等生物学软件进行基因序列及亲缘进化分析。同时收集病例的临床及流行病学信息，采用SPSS软件进行统计分析。结果15份标本来源于12例临床诊断为病毒性脑炎或手足口合并神经症状的患者，8例有临床信息记载的都表现有发热、皮疹、脑膜刺激征和颈项强直的症状，临床症状和发病时间不呈现典型的聚集性。01111、01169和01130三例患者分别采集了两份标本，除了01169来源的两份标本间有一个碱基的差别，两份标本来源的VP1序列核苷酸和氨基酸相似性均为100%。15株毒株之间的核苷酸同源性为94.5%～100%，氨基酸同源性为98.0%～100%。15株毒株和原型株G10的核苷酸同源性为68.5%～70.5%，氨基酸同源性为90.5%～91.9%。系统进化分析显示，本研究15株序列全部属于B1b簇，又分属三小簇。结论引起神经症状的15株CA16均为B1b基因型，深入开展CA16分子流行病学对我国HFMD的防控及疫苗的研发将提供重要参考。
Objective To analyze the VP1 sequences of coxsackievirus A16 （CA16） causing neurologic complications.MethodsClinical samples and epidemiological information were collected from patients with viral encephalitis. Coxsackievirus A16 in these samples were first detected with real time RT-PCR and then isolated. RT-PCR was performed to amplify VP1 sequences and the amplified products were sequenced. DNAStar 5.0 and Mega 5 were used for sequence analysis. All data was analyzed with SPSS statistical software.ResultsFifteen samples were collected from 12 patients with hand, foot and mouth disease（HFMD） complicated by neurologic complications. Eight patients had the symptoms of fever, skin rash, signs of meningeal irritation and neck rigidity. No typical cluster was associated with clinical features or the time of onset. Both pharyngeal/anal swab and serum samples were collected from three patients （patient′s number： 01111, 01169 and 01130）. The two samples collected from both 01111 and 01130 patients shared 100% similarity in nucleotide and amino acid based on VP1 sequences, while those from 01169 patient differed in only one base. The 15 CA16 isolates were highly similar in VP1 gene, sharing 94.5%-100% homology in nucleotide sequences and 98.0%-100% homology in amino acid sequences. These 15 isolates showed 68.5%-70.5% identities in nucleotide sequences and 90.5%-91.9% identities in amino acid sequences with the CA16 prototype strain G10. Phylogenetic analysis revealed that based upon VP1 sequences, all of the 15 CA16 isolates grouped into genotype B subtype 1b （B1b）, which was further classified into three clusters.ConclusionAll of the 15 CA16 isolates causing neurologic complications belonged to B1b sub-genotype. Understanding the molecular epidemiology of CA16 would be essential for controlling morbidity rates of HFMD and vaccine research.
作者
马红霞
潘静静
李懿
卫海燕
杜燕华
李幸乐
黄学勇
许汴利
Ma Hongxia;Pan Jingjing;Li Yi;Wei Haiyan;Du Yanhua;Li Xingle;Huang Xueyong;Xu Bianli(Henan Key Laboratory of Pathogenic Organism, Henan Center for Disease Control and Prevention, Zhengzhou 450016, China)
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2018年第4期274-279,共6页
Chinese Journal of Microbiology and Immunology
基金
河南省基础与前沿技术研究计划项目（142300410077）
河南省医学科技攻关计划省部共建项目（201601028）
河南省卫生科技领军人才
河南省自然科学基金（182300410384）
