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目的分析幼童手足口病病毒EV71衣壳蛋白VP1基因特征,为手足口病临床预防和控制提供指导。方法随机收集604例手足口病幼童患者标本,培养病毒株并分离鉴定。采用RT-PCR扩增EV71及其VP1区基因,经基因产物测序分析毒株的核苷酸和氨基酸序列同源性。结果604例手足口病幼童患者标本中,男性患者369例(61.09%)和女性235例(38.91%);(34.60%).1~岁179例(29.64%),2~岁216例(35.76%)和3~岁209例(34.60%)。共分离出病毒129株,分离率21.36%,其中EV71 61株(占47.29%),CoxA16 28株(占21.71%),其他肠道病毒40株(占31.01%)。61株EV71的VP1区基因序列间核苷酸同源性95.7%~100.0%,氨基酸同源性97.3%~100.0%。与C4a亚型代表株的核苷酸同源性96.4%~98.9%,氨基酸同源性99.0%~100.0%。与CoxA16原始株G-10、A、B基因型代表株VP1区以及C4b、C5亚型代表株的核苷酸同源性分别为66.0%~68.9%、78.6%~82.8%、83.2%~85.5%、95.2%~97.4%和84.5%~87.6%,氨基酸同源性分别为71.4%~75.7%、90.5%~93.6%、95.1%~96.0%、99.0%~99.8%和97.6%~99.4%。结论手足口病发病以3~4岁男性幼童为主,且以EV71为主。所分离的EV71流行株属于C4亚型。
Objective To analyze the characteristics of capsid protein VP1 of hand-foot-mouth disease virus (EV71) in young children, and provide guidance for clinical prevention and control of hand-foot-mouth disease. Methods A total of 604 cases of hand, foot and mouth disease were collected from young children. The virus strains were isolated and identified. The EV71 and its VP1 region genes were amplified by RT-PCR, and the nucleotide and amino acid sequence homologies of the strains were analyzed by gene product sequencing. Results Among the 604 patients with hand-foot-mouth disease, 369 (61.09%) were males and 235 (38.91%) were females (34.60%), 179 (29.64% 35.76%) and 209 cases (34.60%) aged 3 ~ A total of 129 strains of viruses were isolated with a separation rate of 21.36%, of which 61 were EV71 (47.29%), 28 were CoxA16 (21.71%) and 40 were other enteroviruses (31.01%). The nucleotide sequence of the VP1 region of 61 EV71 strains shared 95.7% -100.0% nucleotide homology and 97.3% -100.0% amino acid homology. The nucleotide homology with the C4a subtype was 96.4% ~ 98.9% and the amino acid homology was 99.0% ~ 100.0%. The nucleotide homologies with the representative strains of VP1 and C4b and C5 subtypes of CoxA16 original strain G-10, A and B genotypes were 66.0% -68.9%, 78.6% -82.8% and 83.2% -85.5 %, 95.2% -97.4% and 84.5% -87.6%, respectively. The amino acid homologies were 71.4% -75.7%, 90.5% -93.6%, 95.1% -96.0%, 99.0% -99.8% and 97.6% -99.4%, respectively. Conclusion The incidence of hand-foot-mouth disease is mainly young male aged 3 to 4 years old, and mainly EV71. The isolated strains of EV71 belonged to the C4 subtype.