2017年安庆市急性胃肠炎暴发疫情病原体诺如病毒的鉴定及基因特征分析
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摘要
目的对安徽省安庆市2017年2-3月间发生的10起急性胃肠炎暴发疫情病原诺和病毒进行鉴定及基因分型。方法采集聚集性和暴发疫情中有胃肠炎症状者的肛拭子标本,采用实时荧光定量RT-PCR检测诺如病毒(NoV)核酸,使用RT-PCR对核酸阳性标本扩增NoV衣壳区-聚合酶区,选取27份琼脂糖电泳条带清晰的阳性标本进行基因测序及型别鉴定,并与国内外参考株序列构建进化树,进行遗传进化分析。结果 10起胃肠炎暴发疫情中共采集103份肛拭子标本,实时荧光定量RT-PCR检测诺如病毒核酸阳性41份,且均为GII型,阳性率为39.81%。选取27份阳性标本测序,其中20份测序成功,进化分析显示19条序列为NoV GII.P16/GII.2,1条序列为NoV GII.P17/GII.17。结论诺如病毒GII.P16/GII.2是安庆市2017年2-3月份急性胃肠炎暴发疫情的主要病原体。
Objectives To identify and characterize the pathogen causing 10 outbreaks of gastroenteritis in the City of Anqing between February and March 2017 and to genotype human norovirus. Methods Rectal swabs were collected from patients with symptoms of gastroenteritis during outbreaks.Noroviruses(NoV)in those samples were detected using a real-time reverse transcription-polymerase chain reaction(RT-PCR).The NoV capsid region and the RNA-dependent RNA polymerase(RdRp)region from NoV-positive specimens were amplified using RT-PCR.Twenty-seven positive samples with clear bands in agarose gel electrophoresis were selected for gene sequencing and genotyping.The sequences of the capsid and RdRp regions were analyzed,phylogenetic trees were constructed and NoV was compared to reference strains,and genetic evolutionary analysis was performed. Results One hundred and four rectal swabs were collected during 10 outbreaks of gastroenteritis.NoV was detected in 41(39.81%)of 103 samples according to RT-PCR.All of the isolates were the GII genotype.NoV was successfully sequenced from 20 of 27 positive samples.Phylogenetic tree analysis indicated that most(95%,19/20)of the sequences were from a recombinant GII.P16/GII.2 strain except 1,which was GII.P17/GII.17. Conclusion NoV GII.P16/GII.2 was the leading cause of outbreaks of acute gastroenteritis between February and March 2017 in the City of Anqing.II.P16/GII.2 except one,which was GII.P17/GII.17.Conclusion Norovirus GII.P16/GII.2 was the leading cause of the outbreak of acute gastroenteritis between February and March 2017 in Anqing city.
Objectives To identify and characterize the pathogen causing 10 outbreaks of gastroenteritis in the City of Anqing between February and March 2017 and to genotype human norovirus. Methods Rectal swabs were collected from patients with symptoms of gastroenteritis during outbreaks.Noroviruses(NoV)in those samples were detected using a real-time reverse transcription-polymerase chain reaction(RT-PCR).The NoV capsid region and the RNA-dependent RNA polymerase(RdRp)region from NoV-positive specimens were amplified using RT-PCR.Twenty-seven positive samples with clear bands in agarose gel electrophoresis were selected for gene sequencing and genotyping.The sequences of the capsid and RdRp regions were analyzed,phylogenetic trees were constructed and NoV was compared to reference strains,and genetic evolutionary analysis was performed. Results One hundred and four rectal swabs were collected during 10 outbreaks of gastroenteritis.NoV was detected in 41(39.81%)of 103 samples according to RT-PCR.All of the isolates were the GII genotype.NoV was successfully sequenced from 20 of 27 positive samples.Phylogenetic tree analysis indicated that most(95%,19/20)of the sequences were from a recombinant GII.P16/GII.2 strain except 1,which was GII.P17/GII.17. Conclusion NoV GII.P16/GII.2 was the leading cause of outbreaks of acute gastroenteritis between February and March 2017 in the City of Anqing.II.P16/GII.2 except one,which was GII.P17/GII.17.Conclusion Norovirus GII.P16/GII.2 was the leading cause of the outbreak of acute gastroenteritis between February and March 2017 in Anqing city.
引文
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[18]吴冰珊,黄枝妙,欧剑鸣,等.诺如病毒新重组株GII.P16/GII.2引起福建省2016年冬季病毒性胃肠炎暴发[J].中国人兽共患学报,2017,33(9):805-13.
[19]刘敏鸿,曹孟婵,金爱武,等.诺如病毒GII.17和GI.3基因型引起急性胃肠炎的病原分析[J].安徽省预防医学杂志,2017,23(1):6-8.
[2]Caul E.Small round structured viruses:airborne transmission and hospital control[J].Lancet,1994,343(8908):1240-2.
[3]Fankhauser RL,Monroe SS,Noel JS,et al.Epidemiologic and molecular trends of"Norwalk-like viruses"associated with outbreaks of gastroenteritis in the United States[J].J Infect Dis,2002,186(1):1-7.
[4]Xi JN,Graham DY,Wang KN,et al.Norwalk virus genome cloning and characterization[J].Science,1990,250(4987):1580-3.
[5]Bertolotti-Ciarlet A,Crawford SE,Hutson AM,et al.The 3'end of Norwalk virus mRNA contains determinants that regulate the expression and stability of the viral capsid protein VP1:a novel function for the VP2protein[J].J Virol,2003,77(21):11603-15.
[6]Mesquita JR,Barclay L,Nascimento MS,et al.Novel norovirus in dogs with diarrhea[J].Emerg Infect Dis,2010,16(6):980-2.
[7]Zheng DP,Ando T,Fankhauser RL,et al.Norovirus classification and proposed strain nomenclature[J].Virology,2006,346(2):312-23.
[8]Lopman B,Vennema H,Kohli E,et al.Increase in viral gastroenteritis outbreaks in Europe and epidemic spread of new norovirus variant[J].Lancet,2004,363(9410):682-8.
[9]Bull RA,Tu ET,McIver CJ,et al.Emergence of a new norovirus genotype II.4variant associated with global outbreaks of gastroenteritis[J].J Clin Microbiol,2006,44(2):327-33.
[10]Eden JS,Bull RA,Tu E,et al.Norovirus GII.4variant 2006b caused epidemics of acute gastroenteritis in Australia during 2007and 2008[J].J Clin Virol,2010,49(4):265-71.
[11]Yen C,Wikswo ME,Lopman BA,et al.Impact of an emergent norovirus variant in 2009on norovirus outbreak activity in the United States[J].Clin Infect Dis,2011,53(6):568-71.
[12]Lu J,Sun L,Fang L,et al.Gastroenteritis Outbreaks Caused by Norovirus GII.17,Guangdong Province,China,2014-2015[J].Emerg Infect Dis,2015,21(7):1240-2.
[13]Fu J,Ai J,Jin M,et al.Emergence of a new GII.17norovirus variant in patients with acute gastroenteritis in Jiangsu,China,September 2014to March 2015[J].Euro Surveill,2015,20(24):21157.
[14]Niendorf S,Jacobsen S,Faber M,et al.Steep rise in norovirus cases and emergence of a new recombinant strain GII.P16-GII.2,Germany,winter 2016[J].Euro Surveill,2017,22(4):447.
[15]Lu J,Fang L,Sun L,et al.Association of GII.P16-GII.2Recombinant Norovirus Strain with Increased Norovirus Outbreaks,Guangdong,China,2016[J].Emerg Infect Dis,2017,23(7):1188-90.
[16]Liu LT,Kuo TY,Wu CY,et al.Recombinant GII.P16-GII.2Norovirus,Taiwan,2016[J].Emerg Infect Dis,2017,23(7):1180-3.
[17]冯微宏,肖勇,沙丹,等.2013年无锡地区诺如病毒腹泻病的监测研究[J].中国病原生物学杂志,2015,10(5):430-4.
[18]吴冰珊,黄枝妙,欧剑鸣,等.诺如病毒新重组株GII.P16/GII.2引起福建省2016年冬季病毒性胃肠炎暴发[J].中国人兽共患学报,2017,33(9):805-13.
[19]刘敏鸿,曹孟婵,金爱武,等.诺如病毒GII.17和GI.3基因型引起急性胃肠炎的病原分析[J].安徽省预防医学杂志,2017,23(1):6-8.