西藏自治区埃柯病毒7型和11型基因特征分析
发布时间：2018-09-03 19:40
【摘要】： 人类肠道病毒(human enteroviruses,HEV)属于小RNA病毒科(Picornaviridae),目前已经报道的有101个型,被分为A、B、C、D 4类(species)及脊灰病毒,其中64个血清型被早期的中和试验所定型。肠道病毒基因组全长大约7500nt,VP1区是其重要的蛋白编码区,也是抗原决定区,是其分型的决定区域,近年来新发现的肠道病毒型别均是以VP1区的序列为依据。VP1区核苷酸序列的对比分析也是肠道病毒分子流行病学研究的重要数据资料。 大多数肠道病毒的感染是无症状的或只引起轻微症状,如非特异性发热或轻微上呼吸道症状(普通感冒);也可以引起广泛多样的临床疾病,包括急性出血性结膜炎、无菌性脑膜炎、皮疹、急性弛缓性麻痹、手足口病、心肌炎和新生儿败血症。由于肠道病毒的型别多样,可引起多种轻重疾病,或者形成无症状感染者,近年来越来越受到各国疾病控制部门的重视。 我国地域辽阔,肠道病毒种类多,常年都有疫情发生。西藏自治区地理位置特殊,其肠道病毒的流行特点和型别特征未见相关研究报道。从我们这项研究的结果看,西藏地区1999—2002年人群中分离到polio,ECHO,Cox等多型别肠道病毒,以ECHO7和ECHO11最多,所以我们选取ECHO7和ECHO11病毒做进一步的分子流行病学研究。 目的研究西藏自治区肠道病毒ECHO7和ECHO11 VP1区编码基因特征及其分子流行病学特点。 方法选取从1999年—2002年西藏自治区AFP病例和0—5岁到儿童医院就诊儿童及部分健康儿童的659份粪便标本中分离的16株ECHO7病毒和15株ECHO11病毒,进行RNA提取,VP1编码区RT-PCR,PCR产物的核苷酸序列测定和分析。 结果1999年标本分离到9株ECHO7病毒,2000年分离到7株ECHO7病毒,其它年份未分离到ECHO7病毒。1999年标本分离到15株ECHO11病毒,其它年份未分离到ECHO11病毒。VP1区核苷酸序列测定结果证实这些经血清中和试验定型的病毒确实为ECHO7和ECHO11病毒;16株ECHO7病毒VP1区基因全长都是876bp,翻译的氨基酸全长292aa。所测16株ECHO7病毒核苷酸序列之间同源性在86.73-100.0%之间,所测序列和ECHO7原型株——Wallace株相比,同源性在77.85-78.99%之间;和引起致死性脑脊髓炎的ECHO7变异株——UMMC株相比,同源性在81.67-83.79%之间。本研究构建的ECHO7病毒遗传进化树将已知的ECHO7病毒划分为4个基因型(genotype),西藏分离的ECHO7病毒独自形成1个新基因型(D基因型)。所测15株ECHO11病毒VP1区基因全长都是876bp,翻译的氨基酸全长292aa。序列对比分析和基于VP1全基因序列构建的遗传进化树看,在目前已知的ECHO11病毒形成A、B、C、D 4个基因组(genogroup),A组包括中国山东省大部分分离株,以及中国西藏自治区的4株;B组仅有ECHO11的原型株USA/CA53-Gregory;C组包括中国山东省较特殊的1株,以及中国西藏自治区的11株:15株西藏自治区ECHO11分离株VP1区氨基酸序列对比明显显示出,分入A组和C组西藏毒株在292aa中有19aa的差异。这2组的西藏毒株同源性组间差异显著大于组内差异。 结论首次划分了ECHO7病毒的基因型,首次报告了我国西藏地区1999年和2000年人群中流行的ECHO7病毒为一个新的基因型,D基因型;16株D基因型病毒划分为2个基因亚型,D1和D2。1999年流行的为D1;2000年流行的为D2。西藏地区1999年和2000年ECHO7流行为不同的病毒亚型引起。2001年—2002年未发现ECHO7的持续流行。目前我国分离的ECHO11病毒存在2个基因型,A和C;西藏自治区1999年存在A和C 2个亚型ECHO11病毒的共循环。
[Abstract]:Human enteroviruses (HEV) belong to the family Picornaviridae. There are 101 types of human enteroviruses, which are classified into A, B, C, D 4 and poliovirus. 64 serotypes are typed by early neutralization tests. The whole genome of HEV is about 7,500 nt, and VP1 region is an important protein coding region. The antigenic determinant region is the decisive region for the typing of enteroviruses. The newly discovered enterovirus types in recent years are based on the sequence of VP1 region. The comparative analysis of nucleotide sequences in VP1 region is also an important data for the molecular epidemiology of enteroviruses.
Most enterovirus infections are asymptomatic or cause only mild symptoms, such as nonspecific fever or mild upper respiratory symptoms (common cold); they can also cause a wide variety of clinical diseases, including acute hemorrhagic conjunctivitis, aseptic meningitis, rashes, acute flaccid paralysis, hand-foot-mouth disease, myocarditis and neonatal sepsis. In recent years, more and more attention has been paid to enterovirus by disease control departments in various countries because of its various types, which can cause a variety of serious and mild diseases, or form asymptomatic infections.
Tibet Autonomous Region has a special geographical location, and its epidemic characteristics and types of enteroviruses have not been reported. From the results of our study, polio, ECHO, Cox and other enteroviruses were isolated from the population of Tibet from 1999 to 2002, with ECHO7 and E. CHO11 is the largest, so we chose ECHO7 and ECHO11 viruses for further molecular epidemiological study.
Objective To study the coding gene characteristics and molecular epidemiology of enterovirus ECHO7 and ECHO11 VP1 regions in Tibet Autonomous Region.
Methods 16 strains of ECHO7 virus and 15 strains of ECHO11 virus were isolated from 659 stool specimens of AFP cases in Tibet Autonomous Region and children from 0 to 5 years old to children's hospital and some healthy children from 1999 to 2002. RNA extraction, VP1 coding region RT-PCR and nucleotide sequencing of PCR products were performed.
Results Nine strains of ECHO7 virus were isolated in 1999, seven strains of ECHO7 virus were isolated in 2000, and 15 strains of ECHO11 virus were isolated in 1999, and none in other years. The nucleotide sequence analysis of VP1 region confirmed that these serum-neutralized and tested viruses were indeed ECHO7 and ECHO11 viruses. The nucleotide sequence homology of 16 strains of ECHO7 virus ranged from 86.73 to 100.0%, and the homology between the nucleotide sequence and the prototype strain Wallace strain was 77.85-78.99%. The genetic evolution tree of ECHO7 virus constructed in this study divided the known ECHO7 virus into four genotypes, and the ECHO7 virus isolated from Tibet formed a new genotype (D genotype). The VP1 region of 15 strains of ECHO11 virus were 876 BP in length and 292aa in amino acid translation. According to the analysis and genetic evolution tree based on VP1 gene sequence, four genomes (A, B, C, D) of ECHO11 virus were formed, group A included most isolates from Shandong Province of China, and four isolates from Tibet Autonomous Region of China; group B only had the prototype strain USA/CA53-Gregory of ECHO11; group C included a more special strain from Shandong Province of China. Compared with 11 strains from Tibet Autonomous Region of China: 15 strains from ECHO11 of Tibet Autonomous Region, the amino acid sequence of VP1 region showed that there were 19 AA differences in 192aa among the Tibetan strains divided into group A and group C.
Conclusion The genotypes of ECHO7 virus were firstly classified, and a new genotype, D genotype, was reported for the first time in Tibet in 1999 and 2000. Sixteen strains of D genotype virus were classified into two genotypes, D1 and D2.1999 were prevalent as D1, and D2.1999 and 2000 were prevalent as ECHO7 in Tibet. No persistent epidemic of ECHO7 was found between 2001 and 2002. At present, two genotypes, A and C, of the ECHO11 virus isolated in China, and a co-circulation of two subtypes, A and C, of the ECHO11 virus were found in Tibet Autonomous Region in 1999.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R373
本文编号：2220974
[Abstract]:Human enteroviruses (HEV) belong to the family Picornaviridae. There are 101 types of human enteroviruses, which are classified into A, B, C, D 4 and poliovirus. 64 serotypes are typed by early neutralization tests. The whole genome of HEV is about 7,500 nt, and VP1 region is an important protein coding region. The antigenic determinant region is the decisive region for the typing of enteroviruses. The newly discovered enterovirus types in recent years are based on the sequence of VP1 region. The comparative analysis of nucleotide sequences in VP1 region is also an important data for the molecular epidemiology of enteroviruses.
Most enterovirus infections are asymptomatic or cause only mild symptoms, such as nonspecific fever or mild upper respiratory symptoms (common cold); they can also cause a wide variety of clinical diseases, including acute hemorrhagic conjunctivitis, aseptic meningitis, rashes, acute flaccid paralysis, hand-foot-mouth disease, myocarditis and neonatal sepsis. In recent years, more and more attention has been paid to enterovirus by disease control departments in various countries because of its various types, which can cause a variety of serious and mild diseases, or form asymptomatic infections.
Tibet Autonomous Region has a special geographical location, and its epidemic characteristics and types of enteroviruses have not been reported. From the results of our study, polio, ECHO, Cox and other enteroviruses were isolated from the population of Tibet from 1999 to 2002, with ECHO7 and E. CHO11 is the largest, so we chose ECHO7 and ECHO11 viruses for further molecular epidemiological study.
Objective To study the coding gene characteristics and molecular epidemiology of enterovirus ECHO7 and ECHO11 VP1 regions in Tibet Autonomous Region.
Methods 16 strains of ECHO7 virus and 15 strains of ECHO11 virus were isolated from 659 stool specimens of AFP cases in Tibet Autonomous Region and children from 0 to 5 years old to children's hospital and some healthy children from 1999 to 2002. RNA extraction, VP1 coding region RT-PCR and nucleotide sequencing of PCR products were performed.
Results Nine strains of ECHO7 virus were isolated in 1999, seven strains of ECHO7 virus were isolated in 2000, and 15 strains of ECHO11 virus were isolated in 1999, and none in other years. The nucleotide sequence analysis of VP1 region confirmed that these serum-neutralized and tested viruses were indeed ECHO7 and ECHO11 viruses. The nucleotide sequence homology of 16 strains of ECHO7 virus ranged from 86.73 to 100.0%, and the homology between the nucleotide sequence and the prototype strain Wallace strain was 77.85-78.99%. The genetic evolution tree of ECHO7 virus constructed in this study divided the known ECHO7 virus into four genotypes, and the ECHO7 virus isolated from Tibet formed a new genotype (D genotype). The VP1 region of 15 strains of ECHO11 virus were 876 BP in length and 292aa in amino acid translation. According to the analysis and genetic evolution tree based on VP1 gene sequence, four genomes (A, B, C, D) of ECHO11 virus were formed, group A included most isolates from Shandong Province of China, and four isolates from Tibet Autonomous Region of China; group B only had the prototype strain USA/CA53-Gregory of ECHO11; group C included a more special strain from Shandong Province of China. Compared with 11 strains from Tibet Autonomous Region of China: 15 strains from ECHO11 of Tibet Autonomous Region, the amino acid sequence of VP1 region showed that there were 19 AA differences in 192aa among the Tibetan strains divided into group A and group C.
Conclusion The genotypes of ECHO7 virus were firstly classified, and a new genotype, D genotype, was reported for the first time in Tibet in 1999 and 2000. Sixteen strains of D genotype virus were classified into two genotypes, D1 and D2.1999 were prevalent as D1, and D2.1999 and 2000 were prevalent as ECHO7 in Tibet. No persistent epidemic of ECHO7 was found between 2001 and 2002. At present, two genotypes, A and C, of the ECHO11 virus isolated in China, and a co-circulation of two subtypes, A and C, of the ECHO11 virus were found in Tibet Autonomous Region in 1999.
【学位授予单位】：中国协和医科大学
【学位级别】：硕士
【学位授予年份】：2007
【分类号】：R373
【引证文献】
相关硕士学位论文 前1条
1 晁灵;2008年河南省埃柯病毒25型基因特征分析[D];郑州大学;2011年
，本文编号：2220974
本文链接：https://www.wllwen.com/yixuelunwen/binglixuelunwen/2220974.html
最近更新
教材专著
