2009-2015年福建省福州市急性腹泻儿童中诺如病毒基因多样性及动态变化研究

Genetic diversity and its dynamic variation of noroviruses causing acute diarrhea in children in Fuzhou,2009-2015

目的通过定点监测了解2009-2015年福建省福州市急性腹泻儿童中诺如病毒基因多样性及动态变化趋势。方法采集2009-2015年福州市儿童医院因急性腹泻入院的5岁以下儿童的粪便标本,应用实时荧光定量聚合酶链反应(Real-time PCR)筛查诺如病毒阳性标本。应用反转录聚合酶链反应(RT-PCR)方法扩增完整的VP1区和部分RdRp区并测序,应用在线基因分型工具和种系进化分析方法了解福州市5岁以下儿童中诺如病毒基因型/亚型的分布。结果在随机抽取的93份标本中,Real-time PCR法检测阳性有91份;共获得其中73份完整VP1区和72份部分RdRp区基因序列。在完整的VP1分型中,GⅡ.4_2006b型43份,GⅡ.4 Syndeny_2012型16份,GⅡ.3型10份,GⅡ.4 New Orleans_2009型2份,GⅡ.12型1份,GⅡ.7型1份。在部分RdRp区分型中,共发现GⅡ.P4_2006b型43份,GⅡ.Pe型16份,GⅡ.P12型10份,GⅠ.P4型1份,GⅠ.Pa型1份,GⅡ.P21型1份。结论应用双命名系统分型的方法,发现2009-2015年福州市急性腹泻儿童中诺如病毒具有遗传多样性,其中2009-2012年流行的优势毒株为GⅡ.P4_2006b/GⅡ.4_2006b型,2013-2015年流行的优势毒株为GⅡ.Pe/GⅡ.4 Sydney_2012型。此外,GⅡ.P12/GⅡ.3为福州市持续流行的型别。

Objective To investigate the genetic diversity and explore the dynamic variation of genotypes of noroviruses causing acute diarrhea in children in Fuzhou.Methods Fecal specimens were collected from sick children aged 〈5 years hospitalized in Fuzhou Children Hospital due to acute diarrhea from 2009 to 2015.Real-time fluorescence quantitativepolymerase chain reaction（ Real-time PCR） was performed to identify the norovirus infection.For the positive specimens,complete viral VP1 regions and partial RdRp regions were amplified by（ reverse transcription-ploymerase chain reaction）（ RTPCR） and sequenced.The online genotyping tools and phylogenetic analysis methods were used to genotype the noroviruses associated with acute diarrhea in children aged 5 years in Fuzhou.Results In 93 specimens randomly selected,91 were noroviruses positive in real-time PCR,complete VP1 from 73 noroviruses strains and partial RdRp region from 72 noroviruses strains were obtained.Complete VP1-based online genotyping revealed that 43 strains belonged to GⅡ.4_2006b,16 belonged to GⅡ.4 Syndeny_2012,10 belonged to GⅡ.3 and 2 belonged to GⅡ.4 NewOrleans_2009,and 1 belonged to GⅡ.12 and GⅡ.7 respectively.Partial RdRp-based online genotyping showed that 43 strains belonged to GⅡ.P4_2006b,16 belonged to GⅡ.Pe,10 belonged to GⅡ.P12 and 1 belonged to GⅠ.P4,GⅠ.Pa and GⅡ.P21 respectively.Conclusion Combinatorial nomenclature of complete VP1 and partial RdRp regions analysis revealed that the noroviruses causing acute diarrhea in children aged 〈5 years in Fuzhou were geneticallydiverse during 2009-2015.G Ⅱ.P4_ 2006b/G Ⅱ.4_ 2006 b was predominant during 2009-2012 and G Ⅱ.Pe/G Ⅱ.4 Syndeny_2012 was predominant during 2013-2015.Besides,G Ⅱ.P12/GⅡ.3 was the type detected consecutively in Fuzhou.