2013-2014年漳州市手足口病的病原学特征及分子流行病学研究

许美燕; 蔡茂荣; 胡慧玲; 林淑银

doi:10.3784/j.issn.1003-9961.2016.10.007

2013-2014年漳州市手足口病的病原学特征及分子流行病学研究

Etiology and molecular epidemiology of hand, foot and mouth disease in Zhangzhou, 2013-2014

摘要

摘要: 目的了解2013-2014年漳州市手足口病的病原构成及分子流行病学特征，为漳州市手足口病的防治工作提供科学依据。方法采用实时荧光定量反转录聚合酶链反应（Real-time RT-PCR）方法，检测2013-2014年漳州市哨点医院送检的1421份临床诊断手足口病疑似病例的标本，进行肠道病毒通用型、肠道病毒71型（EV71）和柯萨奇病毒A组16型（Cox A16）的核酸检测；通过RT-PCR方法，扩增VP1区基因部分片段并测序，对非EV71及非Cox A16的肠道病毒进行分型鉴定；扩增完整VP1区基因片段并测序，构建系统进化树对EV71、Cox A16、Cox A6和Cox A10进行基因进化分析。结果 1063份标本肠道病毒通用型阳性，阳性率为74.81%，其中222份标本EV71阳性，340份Cox A16阳性，8份EV71+Cox A16阳性，493份非EV71及非Cox A16的肠道病毒阳性。随机抽取40份非EV71及非Cox A16的肠道病毒阳性标本经RT-PCR和测序进一步分型，共鉴定出38份非EV71及非Cox A16的肠道病毒，其中Cox A6型25份（65.79%，25/38），Cox A10型8份（21.05%，8/38），Cox A5型1份（2.63%，1/38），Cox A4型3份（7.89%，3/38）和Cox A9型1份（2.63%，1/38）。VP1区完整基因的进化分析显示，漳州市的EV71均属C4基因亚型的C4a进化分支，Cox A16分属B1基因亚型的B1a和B1b两个分支；Cox A6和Cox A10均与原型株及国外地区分离株的亲缘关系远，与国内地区分离株的亲缘关系近。结论 2013-2014年漳州市非EV71及非Cox A16的肠道病毒在手足口病病原谱中占有重要位置；非EV71及非Cox A16的肠道病毒型别多样，Cox A6和Cox A10为其中的优势型别。漳州市的EV71、Cox A16、Cox A6和Cox A10标本在进化树上与国内地区相应分离株共进化共循环。

Abstract: Objective To explore the pathogenic spectrum and molecular epidemiology characteristics of hand foot and mouth disease (HFMD) in Zhangzhou, Fujian province, during 2013-2014 and provide evidence for the prevention and treatment of HFMD. Methods During this period, a total of 1421 samples were collected from clinical diagnosed HFMD cases in sentinel hospitals in Zhangzhou for EV71, Cox A16 and other enterovirus detection with real-time reverse transcription polymerase chain reaction (Real-time RT-PCR). The genotyping of non-EV71 and non-Cox A16 enteroviruses through sequencing partial VP1 coding gene were performed by RT-PCR. Full-length coding regions for the VP1 genes of EV71, Cox A16, Cox A6 and Cox A10 were amplified and sequenced to construct phylogenetic trees for phylogenetic analysis. Results Among the samples tested, 1063 were positive for enteroviruses (74.81%), including 222 EV71 positive samples, 340 Cox A16 positive samples, 8 EV71+Cox A16 positive samples and 493 non-EV71or non-Cox A16 enterovirus positive samples. Forty non-EV71 and non-Cox A16 enterovirus positive samples were selected randomly for further analysis by RT-PCR and sequencing, and among the 38 samples with definite results, 25 were Cox A6 positive (65.79%), 8 were Cox A10 positive(21.05%), 3 were Cox A4 positive (7.89%), 1 was Cox A5 positive (2.63%) and 1 was Cox A9 positive (2.63%). The phylogenetic analysis indicated that all the EV71 strains belonged to evolutionary branch C4a of gene subtype C4, and Cox A16 strains belonged to evolutionary branch B1a and B1b of gene subtypes B1. Phylogeneic analysis also indicated that Cox A6 and Cox A10 strains from Zhangzhou shared low homology with prototype strains and foreign strains but high homology with domestic strains. Conclusion Non-EV71 and non-Cox A16 enterovirus accounted for a high proportion in the pathogen spectrum of HFMD in Zhangzhou during 2013-2014. Five serotypes of non-EV71 and non-Cox A16 enteroviruses were successfully identified, and Cox A6 and Cox A10 were predominant. Phylogenetic analysis showed that EV71, Cox A16, Cox A6 and Cox A10 co-circulated and co-evolved with the strains detected both at home and abroad in Zhangzhou.

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