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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。
Objective To understand the etiopathogenisis and molecular epidemiological characteristics of HFMD in Zhangzhou City from 2013 to 2014 so as to provide a scientific basis for prevention and control of HFMD in Zhangzhou City. Methods Real-time RT-PCR was used to detect 1421 suspected cases of HFMD in sentinel hospital of Zhangzhou City from 2013 to 2014, (EV71) and Cox A16 (Cox A16). The partial VP1 gene fragment was amplified by RT-PCR and sequenced. The non-EV71 and non-EV71 Cox A16 and Cox A16. The complete VP1 gene fragment was amplified and sequenced. The phylogenetic tree was constructed to analyze the evolution of EV71, Cox A16, Cox A6 and Cox A10. Results The positive rate of 1063 samples of enterovirus was 74.81%, of which 222 samples were EV71 positive, 340 Cox A16 positive, 8 EV71 + Cox A16 positive, 493 non-EV71 and non-Cox A16 enterovirus Positive. A total of 40 non-EV71 and non-Cox A16 enterovirus viruses were enrolled from 40 non-EV71 and non-Cox A16 positive enterovirus positive samples by RT-PCR and sequencing. Among them, 25 were Cox A6 type (65.79 , Cox A10 type (21.05%, 8/38), Cox A5 type 1 (2.63%, 1/38), Cox A4 type 3 (7.89%, 3/38) and Cox A9 type 1 (2.63%, 1/38). The evolutionary analysis of the complete gene of VP1 showed that EV71 in Zhangzhou belongs to the C4a evolutionary branch of C4 gene subtypes. Cox A16 belonged to B1a and B1b subtypes of B1 gene subtypes. Cox A6 and Cox A10 were both associated with the prototype and The isolates in other regions are far away from each other and are closely related to the isolates in China. Conclusion The non-EV71 and non-Cox A16 enteroviruses in Zhangzhou from 2013 to 2014 occupy an important position in the pathogen spectrum of hand-foot-mouth disease. The non-EV71 and non-Cox A16 enterovirus types are diverse, including Cox A6 and Cox A10 Advantageous type. The EV71, Cox A16, Cox A6 and Cox A10 specimens of Zhangzhou City were evolved and co-circulated with the corresponding isolates in China.