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目的了解广州地区2010年肠道病毒(EV)71病毒流行状况。方法设计肠道病毒71型实时荧光RT-PCR引物和TaqMan探针,运用TaqMan实时荧光RT-PCR技术,对2010年全年来医院就诊的疑似手口足病患儿送检标本20150份,进行肠道病毒71型荧光RT-PCR检测,同时,设计了扩增EV71病毒全基因组的引物,随机选取了5株病毒进行全基因组进行测定。结果在20150份临床标本中,EV71病毒的阳性为4780份,全年总的检出率为23.72%,每个月均有阳性检出;其中,EV71病毒阳性率较高的为4～7月份,最高检出率为6月份,达32.19%,最低检出率为10月份,为7.91%;扩增了5株广州株EV71病毒全基因组全长序列,提交到GenBank上,将5株广州株EV71病毒与EV71的A、B3、B4、B5、C1、C2、C3、C4型及阜阳株全基因组核苷酸序列进行ClustalW比较,发现与C4a型阜阳株等同源性为98.00%～99.00%,与C4b同源性为91.00%～93.00%,与C1-C3同源性为82.00%～83.00%,与B3、B4、B5同源性为81.00%～83.00%,与A型同源性为80.00%。结论 2010年广州地区手足口病的主要病原是肠道病毒71型,EV71病毒在2010年全年还是较普遍流行,全基因组序列分析表明,2010年分离的5株广州株EV71病毒属于C4a型。
Objective To understand the prevalence of enterovirus 71 virus in Guangzhou area in 2010. Methods To design enterovirus 71 real-time fluorescent RT-PCR primers and TaqMan probes, and to use real-time fluorescent TaqMan real-time RT-PCR to test 20,150 specimens of suspected hand-foot-mouth disease in hospitals for the year 2010, The virus 71 type fluorescent RT-PCR was used to detect the whole genome of the EV71 virus. At the same time, five genome-wide primers were selected to amplify the whole genome of the EV71 virus. Results In 20,150 clinical samples, the positive rate of EV71 virus was 4780, the overall positive rate was 23.72%, which was positive every month. Among them, the positive rate of EV71 was 4 ~ 7 months , The highest detection rate was 32.19% in June and the lowest detection rate was 7.91% in October. The full-length full-length sequence of five strains of EV71 in Guangzhou was amplified and submitted to GenBank. Five strains of Guangzhou The ClustalW nucleotide sequences of EV71 and EV71 strains A, B3, B4, B5, C1, C2, C3, C4 and Fuyang were 98.00% -99.00% , The homology with C4b was 91.00% -93.00%, the homology with C1-C3 was 82.00% -83.00%, the homology with B3, B4 and B5 was 81.00% -83.00%, the homology with type A was 80.00%. Conclusion The main pathogen of hand-foot-mouth disease in Guangzhou in 2010 was enterovirus 71, and the EV71 virus was prevalent in 2010. Genome-wide sequence analysis showed that 5 strains of EV71 were isolated from Guangzhou in 2004 and belong to type C4a.