2015-2016年贵州省手足口病非EV71、非CVA16肠道病毒型别鉴定和分析

目的 探讨2015-2016年贵州省手足口病非EV71(Enterovirus 71,EV 71)、非CVA16(Coxsackievirus A16,CVA16)肠道病毒株的病原构成及优势型别。方法 共收集2015-2016年贵州省儿童手足口病疑似患者肛拭子350例标本,应用实时荧光定量PCR法筛选出非EV71、非CVA16型肠道病毒,采用 RT-PCR法扩增病毒VP1区序列,PCR阳性扩增产物进行测序,对测得序列进行型别鉴定并与国内外各型代表株进行核苷酸同源性分析,构建系统进化树。结果 从收集到的手足口病疑似患者肛拭子标本中,共分离到21株其他肠道病毒,分别为:CVA5、CVA7、CVA9、CVB3(Coxsackievirus B3,CVB3)、 CVB4 、E11(Echovirus11,ECHO11)、 E17各1株,CVA10 、CVB1各2株,CVB5、E7各5株。通过系统发育树揭示,分离到的同型别毒株之间具有同源性,而非同型别的毒株与国内外各代表株之间具有同源性。结论 贵州省引起手足口病的非EV71、非CVA16型肠道病毒型别分布较广,包括CVB组,CVA组和ECHO病毒,其中CVB5、E7为非EV71、非CVA16型肠道病毒主要病原体。

Isolation and identification on non-EV71,non-CV-A16 enterovirus of hand-foot-mouth disease in Guizhou,2015-2016

Objective To explore the pathogenic constitution and predominant types of Enterovirus 71 (EV 71) and non-CVA16 (CVA16) enterovirus strains of hand,foot and mouth disease (HFMD) in Guizhou province during 2015-2016. Methods A total of 350 samples of rectal swabs from children suspected of HFMD were collected from 2015 to 2016 in Guizhou province to screen non-EV71 and non-CVA16 enterovirus strains by real-time fluorescence quantitative PCR.The viral VP1 region was amplified by RT-PCR and the PCR positive amplification products were sequenced.The tested sequence was identified,and nucleotide homology analysis was performed with all types of representative strains at home and abroad to construct the phylogenetic tree. Results A total of 21 strains of other types of enteroviruses were isolated from swab specimens of suspected HFMD patients which were one strain of CVA5,CVA7,CVA9,CVB3 (Coxsackievirus B3),CVB4,E11 (ECHO11) and E17 respectively,two strains of CVA10 and CVB1 respectively and five strains of CVB5 and E7 respectively.The homology showed between the isolates of the same type and between the representative strains of the different types at home and abroad. Conclusion The non-EV71 and non-CVA16 enterovirus,which cause HFMD in Guizhou province,are widely distributed,including CVB group,CVA group and ECHO virus,among which CVB5 and E7 are the main pathogens.