北京地区成人腹泻诺如病毒的流行和分子特征的初步研究
发布时间：2018-09-05 16:11
【摘要】： 诺如病毒(Noroviruses,NoV)属杯状病毒科诺如病毒属,单股正链RNA病毒,与札如病毒(Sappoviruses,SVs)合称人类杯状病毒,可通过与病人直接、间接接触过的物品、环境而可引起各年龄人群爆发、散发急性非细菌性胃肠炎,临床症状表现为腹泻、腹痛、呕吐、低烧。这两个属具有基因多样性,分5个遗传组(genogroups):GⅠ到GⅤ。其中GⅠ、GⅡ和GⅣ感染人类,根据衣壳序列多样性GⅠ、GⅡ和GⅣ基因组下分为多于8个、17个、1个基因型。常引起人类腹泻的基因组是NoVGⅡ型,GⅡ-4是近年来引起全球流行和散发的主要基因型,由其变异的新株也迅速取代其它株成为优势株,除此以外,动物身上也检测到GⅡ型,由此人们质疑疾病动物源性 NVs是Kapikian于1972年首次发现,大约有7.5kb基因,包含3个开放阅读框架(ORFs)。分别编码RNA聚合酶在内的多蛋白前体、衣壳蛋白、功能未知的碱性蛋白。在一些杯状病毒中,ORF1、ORF2重叠形成聚合蛋白,许多数据表明每隔1-2年就有新的GⅡ-4诺如病毒变异株出现并引起流行,这些变异株包括95/96US株(1995年-2001年)、Farmingtong Hills株(2002年)、Hunter株(2004年)、Sakai株(2004年)、2006a株和2006b株(2006年) NVS分为五个基因组GⅠ-GⅤ,每个基因组包含许多基因型,通常根据衣壳区部分或全部序列进行基因分组、分型,根据Green et al,和Ando et al。的分类:GⅠ,GⅡ,GⅢ由7、8、2个基因型组成,GⅣ,GⅤ各有一个基因型,这种分类随着新株的出现不段更新着。Kageyama提出分成31个基因型,14个GⅠ、17个GⅡ;zheng等人提出8个GⅠ、17个GⅡ;针对分类法许多人提出不同的建议,因为这么多的遗传差异可直接导致聚合酶区重组及株重组,重组株的出现对传统的分类法提出挑战,重组可以定义为序列不同的聚合酶区、衣壳区互相组合产生的新的基因型。因为聚合酶区与衣壳区分属不同型,单独通过衣壳区序列分型代表整个病毒分型显然不适合重组株的命名,另外重组点多位于ORF1/2重叠区,单纯衣壳区的序列决定组、型可丢失重组的判别 对于RNA病毒,重组是一件很平常的事,RNA病毒通过交换序列产生新的基因组、是产生新重组株的动力,是RNA病毒进化的重要机制。RNA病毒在复制过程中由于缺乏校正活性基因组经常发生重组、断裂使得病毒具有高变异率。在逆转录病毒,虫媒病毒,肠道病毒,植物病毒中都有报道,但是在负链RNA病毒中报道甚少。正链病毒含有GORS结构,在RNA病毒复制和进化过程中发挥重要作用。 目前已有多株NoVs重组株报道。来自美国Norfolk的Arg320 RdRp区与Lordsdale株(GⅡ.4)相似,衣壳区与Mexican(GⅡ.3)株相似,泰国的Mc37株ORF1序列与Saitama U1株有97.2%的同源性但是在ORF2和ORF3仅有71.3%和67.9%的同源性。MD145-12株是Lordsdale、Gifu96、SaitamaU1、U3、U4、U16、U17 and U25的重组。GⅡb也是一株重组株,RNA聚合酶区与现已知道的任一基因型不符合,衣壳区与Hawaii,Toronto,Snow Mountain这三个基因型接近,但是病毒学方法肯定的因重组引起的腹泻爆发还很少。 为较好了解我国2007年-2008年秋冬季节诺如病毒的分子流行病学特点,诺如病毒重组株的分子特征,我们开展了本项研究。完成了对北京地区一家三甲综合医院15岁以上成人2007年9月至2008年2月间547份急性胃肠炎病毒病原的筛查。首先采用ELISA法初筛A组轮状病毒,随后用RT-PCR法或PCR法对2007年9月-10月332份标本进行了9种胃肠炎相关病毒(诺如病毒、札如病毒、初筛阳性A组轮状病毒、轮状病毒(B、C)、腺病毒、小双节RNA病毒、曲状病毒、冠状病毒)的检测,对2007年11月-2008年2月的215份标本只检测了诺如病毒。发现在2007年9月-10月332份标本中诺如病毒阳性标本41份,阳性率为12.3%,腺病毒14份(4.2%)、小双节RNA病毒4份(1.5%),A组轮状病毒2份(0.6%),B、C族轮状病毒、曲状病毒、冠状病毒为零,诺如病毒阳性率高于其他8种病毒,是引起成人腹泻主要病毒病原。在总计547份腹泻标本中,106例腹泻是由NoV病毒引起,3例由SVs引起。106株诺如病毒毒株中,96株属于GⅡ,10株属于GⅠ,百分率分别为90.6%和9.4%。进化分析发现2007-2008年秋冬季北京地区共有10种遗传型NoV病毒循环,分别为GⅡ-4,GⅠ-3,GⅠ-2,GⅡ-16、GⅠ-4、GⅡ-d、GⅡ-b和3种不属于任何已知型别的诺如病毒。GⅡ-4为最常见的基因型,90例,发病率是16.4%(90/547),引起散发腹泻的基因型与曾经引起世界范围爆发的代表株相近。在15-24、25-34、35-44、45-60、60岁5个年龄组中,NoV病毒阳性率分别为16.3%、15.8%、27.2%、19.1%、20.6%。35-44年龄组的诺如病毒发病率(27.2%)高于其他年龄组,但年龄性别之间没有统计学差异。时间发病高峰为1月份。很意外的是我们随机挑选聚合酶区同源性较高的标本做外壳区的扩增,了解变异性的可能,发现重组现象多见,有GⅠ-3/GⅠ-8、GⅠ-2/GⅠ-6、GⅡ-6/GⅡ-14、GⅡ-4/GⅡ-3、GⅡ-b/GⅡ-13、GⅡ-d/GⅡ-18等几种重组病毒,认为病毒重组现象普遍,是病毒适应环境自我进化的本能,也是引起腹泻不可忽视的原因。
[Abstract]:Noroviruses (NoV) belong to the genus Noroviruses of the family Caliviridae. Single stranded positive strand RNA viruses, together with Sappoviruses (SVs), are known as human calicivirus viruses. They can cause outbreaks of acute nonbacterial gastroenteritis in people of all ages through direct or indirect contact with patients. The clinical symptoms are diarrhea and abdomen. Pain, vomiting, and low fever. These two genera are genetically diverse and are divided into five genotypes: GI to GV. Among them, GI, GII and GIV infect humans. According to the diversity of capsid sequences, GI, GII and GIV are divided into more than eight, 17, and one genotype. The main genotypes of epidemic and sporadic diseases are rapidly replaced by new strains of mutation. In addition, G type II is also detected in animals, which raises questions about the animal origin of the disease.
NVs were first discovered by Kapikian in 1972. There are about 7.5 KB genes, including three open reading frames (ORFs). They encode multiple protein precursors including RNA polymerases, capsid proteins, and basic proteins with unknown functions. In some goblet viruses, ORF1 and ORF2 overlap to form polymeric proteins. Many data indicate that new GII-4 norovirus disease occurs every 1-2 years. The virus variants, 95/96US (1995-2001), Farmingtong Hills (2002), Hunter (2004), Sakai (2004), 2006a and 2006b (2006), emerged and caused epidemics.
NVS is divided into five genomes, GI-GV. Each genome contains many genotypes, usually grouped according to part or all of the capsid region sequences, and typed according to the Greeet al. and Ado et al. classification: GI, GII, GIII consists of 7, 8, 2 genotypes, GIV, GV each has a genotype, this classification is updated with the emergence of new strains. Kageyama proposed to divide into 31 genotypes, 14 GI, 17 GII; Zheng et al. proposed 8 GI, 17 GII; and proposed different suggestions for many taxonomies, because so many genetic differences can directly lead to polymerase domain recombination and plant recombination. The emergence of recombinant strains challenges traditional taxonomy, recombination can be defined as sequence not. Because the polymerase region and capsid region belong to different genotypes, the capsid region typing alone is not suitable for the name of the recombinant strain. In addition, the recombinant sites are mostly located in the ORF1/2 overlap region, and the sequence determination group of the capsid region alone can lose the recombinant type. Distinguish
For RNA viruses, recombination is a common practice. RNA viruses produce new genomes by exchanging sequences. It is the motive force for producing new recombinant strains and an important mechanism for the evolution of RNA viruses. Arboviruses, enteroviruses, and plant viruses have been reported, but few have been reported in negative-stranded RNA viruses. Positive-stranded viruses contain GORS structures and play an important role in the replication and evolution of RNA viruses.
Several NoVs recombinant strains have been reported. Arg 320 RdRp region from Norfolk is similar to Lordsdale strain (G II.4) and capsid region is similar to Mexican strain (G II.3). The ORF1 sequence of Mc37 strains from Thailand is 97.2% homologous to Saitama U1 strain, but only 71.3% and 67.9% homologous to ORF 2 and ORF3. MD145-12 strains are Lordsdale, Gifu96, SaitamaU1, U3, U3. The recombinant strain of U4, U16, U17 and U25. GIIb is also a recombinant strain. The RNA polymerase region does not correspond to any known genotype. The capsid region is close to Hawaii, Toronto, and Snow Mountain genotypes. However, the virologically confirmed outbreaks of diarrhea caused by recombination are rare.
In order to better understand the molecular epidemiological characteristics of norovirus in autumn and winter of 2007-2008 and the molecular characteristics of the recombinant norovirus strain, we carried out this study. 547 cases of acute gastroenteritis virus pathogens were screened from September 2007 to February 2008 in a third-class general hospital in Beijing. Group A rotavirus was screened by ELISA, and then 9 kinds of gastroenteritis-associated viruses (Norovirus, Zaru virus, Group A rotavirus, B, C, adenovirus, Binodular RNA virus, curvilinear virus, coronavirus) were detected by RT-PCR or PCR in 332 specimens from September to October 2007. Norovirus was detected in 41 specimens of 332 specimens from September to October 2007. The positive rate of Norovirus was 12.3%, adenovirus 14 (4.2%), small binodal RNA virus 4 (1.5%), group A rotavirus 2 (0.6%), B, C rotavirus, curvilinear virus, coronavirus 0. Norovirus positive rate was higher than other 8 viruses. Among 547 diarrhea specimens, 106 were caused by NoV, 3 by SVs, 96 by GII, 10 by GI, and the percentage was 90.6% and 9.4% respectively. GII-4, GI-3, GI-2, GII-16, GI-4, GII-d, GII-b and three non-known Noroviruses were the most common genotypes, with a morbidity of 16.4% (90/547). The genotypes causing sporadic diarrhea were similar to those causing outbreaks worldwide. The positive rate of NoV was 16.3%, 15.8%, 27.2%, 19.1%, 20.6% respectively. The incidence of Norovirus in 35-44 age group (27.2%) was higher than that in other age groups, but there was no statistical difference between age and sex. Recombinant viruses, such as GI-3/GI-8, GI-2/GI-6, GII-6/GII-14, GII-4/GII-3, GII-b/GII-13, GII-d/GII-18, have been found to be common. The recombinant viruses are considered to be an instinct of self-evolution in adapting to the environment and a cause of diarrhea.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R373
本文编号：2224784
[Abstract]:Noroviruses (NoV) belong to the genus Noroviruses of the family Caliviridae. Single stranded positive strand RNA viruses, together with Sappoviruses (SVs), are known as human calicivirus viruses. They can cause outbreaks of acute nonbacterial gastroenteritis in people of all ages through direct or indirect contact with patients. The clinical symptoms are diarrhea and abdomen. Pain, vomiting, and low fever. These two genera are genetically diverse and are divided into five genotypes: GI to GV. Among them, GI, GII and GIV infect humans. According to the diversity of capsid sequences, GI, GII and GIV are divided into more than eight, 17, and one genotype. The main genotypes of epidemic and sporadic diseases are rapidly replaced by new strains of mutation. In addition, G type II is also detected in animals, which raises questions about the animal origin of the disease.
NVs were first discovered by Kapikian in 1972. There are about 7.5 KB genes, including three open reading frames (ORFs). They encode multiple protein precursors including RNA polymerases, capsid proteins, and basic proteins with unknown functions. In some goblet viruses, ORF1 and ORF2 overlap to form polymeric proteins. Many data indicate that new GII-4 norovirus disease occurs every 1-2 years. The virus variants, 95/96US (1995-2001), Farmingtong Hills (2002), Hunter (2004), Sakai (2004), 2006a and 2006b (2006), emerged and caused epidemics.
NVS is divided into five genomes, GI-GV. Each genome contains many genotypes, usually grouped according to part or all of the capsid region sequences, and typed according to the Greeet al. and Ado et al. classification: GI, GII, GIII consists of 7, 8, 2 genotypes, GIV, GV each has a genotype, this classification is updated with the emergence of new strains. Kageyama proposed to divide into 31 genotypes, 14 GI, 17 GII; Zheng et al. proposed 8 GI, 17 GII; and proposed different suggestions for many taxonomies, because so many genetic differences can directly lead to polymerase domain recombination and plant recombination. The emergence of recombinant strains challenges traditional taxonomy, recombination can be defined as sequence not. Because the polymerase region and capsid region belong to different genotypes, the capsid region typing alone is not suitable for the name of the recombinant strain. In addition, the recombinant sites are mostly located in the ORF1/2 overlap region, and the sequence determination group of the capsid region alone can lose the recombinant type. Distinguish
For RNA viruses, recombination is a common practice. RNA viruses produce new genomes by exchanging sequences. It is the motive force for producing new recombinant strains and an important mechanism for the evolution of RNA viruses. Arboviruses, enteroviruses, and plant viruses have been reported, but few have been reported in negative-stranded RNA viruses. Positive-stranded viruses contain GORS structures and play an important role in the replication and evolution of RNA viruses.
Several NoVs recombinant strains have been reported. Arg 320 RdRp region from Norfolk is similar to Lordsdale strain (G II.4) and capsid region is similar to Mexican strain (G II.3). The ORF1 sequence of Mc37 strains from Thailand is 97.2% homologous to Saitama U1 strain, but only 71.3% and 67.9% homologous to ORF 2 and ORF3. MD145-12 strains are Lordsdale, Gifu96, SaitamaU1, U3, U3. The recombinant strain of U4, U16, U17 and U25. GIIb is also a recombinant strain. The RNA polymerase region does not correspond to any known genotype. The capsid region is close to Hawaii, Toronto, and Snow Mountain genotypes. However, the virologically confirmed outbreaks of diarrhea caused by recombination are rare.
In order to better understand the molecular epidemiological characteristics of norovirus in autumn and winter of 2007-2008 and the molecular characteristics of the recombinant norovirus strain, we carried out this study. 547 cases of acute gastroenteritis virus pathogens were screened from September 2007 to February 2008 in a third-class general hospital in Beijing. Group A rotavirus was screened by ELISA, and then 9 kinds of gastroenteritis-associated viruses (Norovirus, Zaru virus, Group A rotavirus, B, C, adenovirus, Binodular RNA virus, curvilinear virus, coronavirus) were detected by RT-PCR or PCR in 332 specimens from September to October 2007. Norovirus was detected in 41 specimens of 332 specimens from September to October 2007. The positive rate of Norovirus was 12.3%, adenovirus 14 (4.2%), small binodal RNA virus 4 (1.5%), group A rotavirus 2 (0.6%), B, C rotavirus, curvilinear virus, coronavirus 0. Norovirus positive rate was higher than other 8 viruses. Among 547 diarrhea specimens, 106 were caused by NoV, 3 by SVs, 96 by GII, 10 by GI, and the percentage was 90.6% and 9.4% respectively. GII-4, GI-3, GI-2, GII-16, GI-4, GII-d, GII-b and three non-known Noroviruses were the most common genotypes, with a morbidity of 16.4% (90/547). The genotypes causing sporadic diarrhea were similar to those causing outbreaks worldwide. The positive rate of NoV was 16.3%, 15.8%, 27.2%, 19.1%, 20.6% respectively. The incidence of Norovirus in 35-44 age group (27.2%) was higher than that in other age groups, but there was no statistical difference between age and sex. Recombinant viruses, such as GI-3/GI-8, GI-2/GI-6, GII-6/GII-14, GII-4/GII-3, GII-b/GII-13, GII-d/GII-18, have been found to be common. The recombinant viruses are considered to be an instinct of self-evolution in adapting to the environment and a cause of diarrhea.
【学位授予单位】：中国疾病预防控制中心
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R373
【引证文献】
相关硕士学位论文 前1条
1 韦玉梅;水环境中三种病毒PMA-PCR方法建立与基于焦磷酸测序技术的细菌多样性研究[D];北京林业大学;2013年
，本文编号：2224784
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