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目的:探讨肠道病毒(HEV)相关小儿病毒性脑炎病原谱及分子流行病学特征,为临床治疗提供依据。方法:选取2010年1月~2012年12月珠海市各监测点中的样本,患儿均疑似患病毒性脑炎。病毒株均采用WTO《病毒试验室操作手册》检测,脑脊液或粪便0.2 ml接种RD细胞,操作步骤按说明书进行。结果:观察的545份样本中,分离到HEV115株,分离率为21.1%。在375份脑脊液样本中,分离HEV 72株,分离率为19.2%;在170份粪便样本中,分离HEV 43株,分离率为25.3%。分离的115株病毒共涉及12个血清型,CVB3-5、E25和E30在近3年中均有检出,且分离株最多,为绝对优势株。9例同时采集脑脊液和粪便样本的患儿中,有7例在两类样本均分离到同一血清型病毒,CVB3和CVB5各2例,两组比较,差异无统计学意义(P>0.05);住院天数比较,差异有统计学意义(P<0.05);PICS评分比较,差异有统计学意义(P<0.05);综合疗效比较,差异有统计学意义(P<0.01)。测序株各血清型内nt和aa同源性分别为79.3%~99.8%和89.1%~99.9%,与各原型株的nt和aa的同源性分别为79.1%~99.90%和88.4%~99.8%。结论:由于病原体的基因重组和变异,易使其致病病原改变,各地医务工作者对于HEV的基因变异应做好高度的准备,以防范各地的病毒性脑炎的暴发。
Objective: To investigate the pathogenic spectrum and molecular epidemiological characteristics of enterovirus (HEV) -related pediatric viral encephalitis and provide the basis for clinical treatment. Methods: From January 2010 to December 2012 Zhuhai samples at various monitoring points, children were suspected of viral encephalitis. Virus strains were detected using the WTO “Virus Lab Operation Manual”, cerebrospinal fluid or stool 0.2 ml RD cells inoculated, procedures according to the instructions. Results: Among 545 samples observed, HEV115 strains were isolated with a separation rate of 21.1%. In 375 samples of cerebrospinal fluid, 72 HEV isolates were isolated with a separation rate of 19.2%. In 170 stool samples, 43 HEV isolates were isolated with a separation rate of 25.3%. The 115 isolates were involved in 12 serotypes. CVB3-5, E25 and E30 were all detected in the past three years, with the largest number of isolates and the absolute dominant strains. Of the 9 children with cerebrospinal fluid and stool samples collected at the same time, 7 cases had the same serotype virus in both groups, 2 cases in each of CVB3 and CVB5. There was no significant difference between the two groups (P> 0.05). There were significant differences in the days of hospitalization between the two groups (P <0.05). There was significant difference in PICS scores between the two groups (P <0.05). There was significant difference between the two groups in comprehensive efficacy (P <0.01). The homologies of nt and aa in each serotype of the sequenced strains ranged from 79.3% to 99.8% and 89.1% to 99.9%, respectively. The homologies of nt and aa to the original strains were 79.1% -99.90% and 88.4% -99.8 %. Conclusion: Due to the genetic recombination and mutation of pathogens, the pathogenic pathogens are easy to change. Medical workers around the world should be highly prepared for genetic variation of HEV in order to prevent the outbreak of viral encephalitis in various places.