北京地區(qū)成人腹瀉諾如病毒的流行和分子特征的初步研究
發(fā)布時(shí)間：2018-09-05 16:11
【摘要】： 諾如病毒(Noroviruses,NoV)屬杯狀病毒科諾如病毒屬,單股正鏈RNA病毒,與札如病毒(Sappoviruses,SVs)合稱人類杯狀病毒,可通過與病人直接、間接接觸過的物品、環(huán)境而可引起各年齡人群爆發(fā)、散發(fā)急性非細(xì)菌性胃腸炎,臨床癥狀表現(xiàn)為腹瀉、腹痛、嘔吐、低燒。這兩個(gè)屬具有基因多樣性,分5個(gè)遺傳組(genogroups):GⅠ到GⅤ。其中GⅠ、GⅡ和GⅣ感染人類,根據(jù)衣殼序列多樣性GⅠ、GⅡ和GⅣ基因組下分為多于8個(gè)、17個(gè)、1個(gè)基因型。常引起人類腹瀉的基因組是NoVGⅡ型,GⅡ-4是近年來引起全球流行和散發(fā)的主要基因型,由其變異的新株也迅速取代其它株成為優(yōu)勢株,除此以外,動(dòng)物身上也檢測到GⅡ型,由此人們質(zhì)疑疾病動(dòng)物源性 NVs是Kapikian于1972年首次發(fā)現(xiàn),大約有7.5kb基因,包含3個(gè)開放閱讀框架(ORFs)。分別編碼RNA聚合酶在內(nèi)的多蛋白前體、衣殼蛋白、功能未知的堿性蛋白。在一些杯狀病毒中,ORF1、ORF2重疊形成聚合蛋白,許多數(shù)據(jù)表明每隔1-2年就有新的GⅡ-4諾如病毒變異株出現(xiàn)并引起流行,這些變異株包括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分為五個(gè)基因組GⅠ-GⅤ,每個(gè)基因組包含許多基因型,通常根據(jù)衣殼區(qū)部分或全部序列進(jìn)行基因分組、分型,根據(jù)Green et al,和Ando et al。的分類:GⅠ,GⅡ,GⅢ由7、8、2個(gè)基因型組成,GⅣ,GⅤ各有一個(gè)基因型,這種分類隨著新株的出現(xiàn)不段更新著。Kageyama提出分成31個(gè)基因型,14個(gè)GⅠ、17個(gè)GⅡ;zheng等人提出8個(gè)GⅠ、17個(gè)GⅡ;針對分類法許多人提出不同的建議,因?yàn)檫@么多的遺傳差異可直接導(dǎo)致聚合酶區(qū)重組及株重組,重組株的出現(xiàn)對傳統(tǒng)的分類法提出挑戰(zhàn),重組可以定義為序列不同的聚合酶區(qū)、衣殼區(qū)互相組合產(chǎn)生的新的基因型。因?yàn)榫酆厦竻^(qū)與衣殼區(qū)分屬不同型,單獨(dú)通過衣殼區(qū)序列分型代表整個(gè)病毒分型顯然不適合重組株的命名,另外重組點(diǎn)多位于ORF1/2重疊區(qū),單純衣殼區(qū)的序列決定組、型可丟失重組的判別 對于RNA病毒,重組是一件很平常的事,RNA病毒通過交換序列產(chǎn)生新的基因組、是產(chǎn)生新重組株的動(dòng)力,是RNA病毒進(jìn)化的重要機(jī)制。RNA病毒在復(fù)制過程中由于缺乏校正活性基因組經(jīng)常發(fā)生重組、斷裂使得病毒具有高變異率。在逆轉(zhuǎn)錄病毒,蟲媒病毒,腸道病毒,植物病毒中都有報(bào)道,但是在負(fù)鏈RNA病毒中報(bào)道甚少。正鏈病毒含有GORS結(jié)構(gòu),在RNA病毒復(fù)制和進(jìn)化過程中發(fā)揮重要作用。 目前已有多株NoVs重組株報(bào)道。來自美國Norfolk的Arg320 RdRp區(qū)與Lordsdale株(GⅡ.4)相似,衣殼區(qū)與Mexican(GⅡ.3)株相似,泰國的Mc37株ORF1序列與Saitama U1株有97.2%的同源性但是在ORF2和ORF3僅有71.3%和67.9%的同源性。MD145-12株是Lordsdale、Gifu96、SaitamaU1、U3、U4、U16、U17 and U25的重組。GⅡb也是一株重組株,RNA聚合酶區(qū)與現(xiàn)已知道的任一基因型不符合,衣殼區(qū)與Hawaii,Toronto,Snow Mountain這三個(gè)基因型接近,但是病毒學(xué)方法肯定的因重組引起的腹瀉爆發(fā)還很少。 為較好了解我國2007年-2008年秋冬季節(jié)諾如病毒的分子流行病學(xué)特點(diǎn),諾如病毒重組株的分子特征,我們開展了本項(xiàng)研究。完成了對北京地區(qū)一家三甲綜合醫(yī)院15歲以上成人2007年9月至2008年2月間547份急性胃腸炎病毒病原的篩查。首先采用ELISA法初篩A組輪狀病毒,隨后用RT-PCR法或PCR法對2007年9月-10月332份標(biāo)本進(jìn)行了9種胃腸炎相關(guān)病毒(諾如病毒、札如病毒、初篩陽性A組輪狀病毒、輪狀病毒(B、C)、腺病毒、小雙節(jié)RNA病毒、曲狀病毒、冠狀病毒)的檢測,對2007年11月-2008年2月的215份標(biāo)本只檢測了諾如病毒。發(fā)現(xiàn)在2007年9月-10月332份標(biāo)本中諾如病毒陽性標(biāo)本41份,陽性率為12.3%,腺病毒14份(4.2%)、小雙節(jié)RNA病毒4份(1.5%),A組輪狀病毒2份(0.6%),B、C族輪狀病毒、曲狀病毒、冠狀病毒為零,諾如病毒陽性率高于其他8種病毒,是引起成人腹瀉主要病毒病原。在總計(jì)547份腹瀉標(biāo)本中,106例腹瀉是由NoV病毒引起,3例由SVs引起。106株諾如病毒毒株中,96株屬于GⅡ,10株屬于GⅠ,百分率分別為90.6%和9.4%。進(jìn)化分析發(fā)現(xiàn)2007-2008年秋冬季北京地區(qū)共有10種遺傳型NoV病毒循環(huán),分別為GⅡ-4,GⅠ-3,GⅠ-2,GⅡ-16、GⅠ-4、GⅡ-d、GⅡ-b和3種不屬于任何已知型別的諾如病毒。GⅡ-4為最常見的基因型,90例,發(fā)病率是16.4%(90/547),引起散發(fā)腹瀉的基因型與曾經(jīng)引起世界范圍爆發(fā)的代表株相近。在15-24、25-34、35-44、45-60、60歲5個(gè)年齡組中,NoV病毒陽性率分別為16.3%、15.8%、27.2%、19.1%、20.6%。35-44年齡組的諾如病毒發(fā)病率(27.2%)高于其他年齡組,但年齡性別之間沒有統(tǒng)計(jì)學(xué)差異。時(shí)間發(fā)病高峰為1月份。很意外的是我們隨機(jī)挑選聚合酶區(qū)同源性較高的標(biāo)本做外殼區(qū)的擴(kuò)增,了解變異性的可能,發(fā)現(xiàn)重組現(xiàn)象多見,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等幾種重組病毒,認(rèn)為病毒重組現(xiàn)象普遍,是病毒適應(yīng)環(huán)境自我進(jìn)化的本能,也是引起腹瀉不可忽視的原因。
[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.
【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R373
本文編號(hào)：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.
【學(xué)位授予單位】：中國疾病預(yù)防控制中心
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R373
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 韋玉梅;水環(huán)境中三種病毒PMA-PCR方法建立與基于焦磷酸測序技術(shù)的細(xì)菌多樣性研究[D];北京林業(yè)大學(xué);2013年
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