西藏自治區(qū)�？虏《�7型和11型基因特征分析
發(fā)布時(shí)間：2018-09-03 19:40
【摘要】： 人類腸道病毒(human enteroviruses,HEV)屬于小RNA病毒科(Picornaviridae),目前已經(jīng)報(bào)道的有101個(gè)型,被分為A、B、C、D 4類(species)及脊灰病毒,其中64個(gè)血清型被早期的中和試驗(yàn)所定型。腸道病毒基因組全長大約7500nt,VP1區(qū)是其重要的蛋白編碼區(qū),也是抗原決定區(qū),是其分型的決定區(qū)域,近年來新發(fā)現(xiàn)的腸道病毒型別均是以VP1區(qū)的序列為依據(jù)。VP1區(qū)核苷酸序列的對(duì)比分析也是腸道病毒分子流行病學(xué)研究的重要數(shù)據(jù)資料。 大多數(shù)腸道病毒的感染是無癥狀的或只引起輕微癥狀,如非特異性發(fā)熱或輕微上呼吸道癥狀(普通感冒);也可以引起廣泛多樣的臨床疾病,包括急性出血性結(jié)膜炎、無菌性腦膜炎、皮疹、急性弛緩性麻痹、手足口病、心肌炎和新生兒敗血癥。由于腸道病毒的型別多樣,可引起多種輕重疾病,或者形成無癥狀感染者,近年來越來越受到各國疾病控制部門的重視。 我國地域遼闊,腸道病毒種類多,常年都有疫情發(fā)生。西藏自治區(qū)地理位置特殊,其腸道病毒的流行特點(diǎn)和型別特征未見相關(guān)研究報(bào)道。從我們這項(xiàng)研究的結(jié)果看,西藏地區(qū)1999—2002年人群中分離到polio,ECHO,Cox等多型別腸道病毒,以ECHO7和ECHO11最多,所以我們選取ECHO7和ECHO11病毒做進(jìn)一步的分子流行病學(xué)研究。 目的研究西藏自治區(qū)腸道病毒ECHO7和ECHO11 VP1區(qū)編碼基因特征及其分子流行病學(xué)特點(diǎn)。 方法選取從1999年—2002年西藏自治區(qū)AFP病例和0—5歲到兒童醫(yī)院就診兒童及部分健康兒童的659份糞便標(biāo)本中分離的16株ECHO7病毒和15株ECHO11病毒,進(jìn)行RNA提取,VP1編碼區(qū)RT-PCR,PCR產(chǎn)物的核苷酸序列測定和分析。 結(jié)果1999年標(biāo)本分離到9株ECHO7病毒,2000年分離到7株ECHO7病毒,其它年份未分離到ECHO7病毒。1999年標(biāo)本分離到15株ECHO11病毒,其它年份未分離到ECHO11病毒。VP1區(qū)核苷酸序列測定結(jié)果證實(shí)這些經(jīng)血清中和試驗(yàn)定型的病毒確實(shí)為ECHO7和ECHO11病毒;16株ECHO7病毒VP1區(qū)基因全長都是876bp,翻譯的氨基酸全長292aa。所測16株ECHO7病毒核苷酸序列之間同源性在86.73-100.0%之間,所測序列和ECHO7原型株——Wallace株相比,同源性在77.85-78.99%之間;和引起致死性腦脊髓炎的ECHO7變異株——UMMC株相比,同源性在81.67-83.79%之間。本研究構(gòu)建的ECHO7病毒遺傳進(jìn)化樹將已知的ECHO7病毒劃分為4個(gè)基因型(genotype),西藏分離的ECHO7病毒獨(dú)自形成1個(gè)新基因型(D基因型)。所測15株ECHO11病毒VP1區(qū)基因全長都是876bp,翻譯的氨基酸全長292aa。序列對(duì)比分析和基于VP1全基因序列構(gòu)建的遺傳進(jìn)化樹看,在目前已知的ECHO11病毒形成A、B、C、D 4個(gè)基因組(genogroup),A組包括中國山東省大部分分離株,以及中國西藏自治區(qū)的4株;B組僅有ECHO11的原型株USA/CA53-Gregory;C組包括中國山東省較特殊的1株,以及中國西藏自治區(qū)的11株:15株西藏自治區(qū)ECHO11分離株VP1區(qū)氨基酸序列對(duì)比明顯顯示出,分入A組和C組西藏毒株在292aa中有19aa的差異。這2組的西藏毒株同源性組間差異顯著大于組內(nèi)差異。 結(jié)論首次劃分了ECHO7病毒的基因型,首次報(bào)告了我國西藏地區(qū)1999年和2000年人群中流行的ECHO7病毒為一個(gè)新的基因型,D基因型;16株D基因型病毒劃分為2個(gè)基因亞型,D1和D2。1999年流行的為D1;2000年流行的為D2。西藏地區(qū)1999年和2000年ECHO7流行為不同的病毒亞型引起。2001年—2002年未發(fā)現(xiàn)ECHO7的持續(xù)流行。目前我國分離的ECHO11病毒存在2個(gè)基因型,A和C;西藏自治區(qū)1999年存在A和C 2個(gè)亞型ECHO11病毒的共循環(huán)。
[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.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R373
本文編號(hào)：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.
【學(xué)位授予單位】：中國協(xié)和醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2007
【分類號(hào)】：R373
【引證文獻(xiàn)】
相關(guān)碩士學(xué)位論文 前1條
1 晁靈;2008年河南省�？虏《�25型基因特征分析[D];鄭州大學(xué);2011年
，本文編號(hào)：2220974
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