摘要
目的分析贵州省狂犬病病毒流行毒株与疫苗毒株的糖蛋白基因（G基因）差异，为狂犬病疫苗研制及有效防控措施的制定提供科学依据。方法以RT-PCR扩增贵州省近几年人和犬狂犬病病毒阳性脑组织标本G基因序列，对扩增产物进行序列测定后采用生物信息学软件对序列与疫苗株G基因序列进行比较分析。结果贵州省狂犬病病毒阳性标本经RT-PCR扩增、测序与拼接后得到8株狂犬病病毒G基因全长序列，与我国常用狂犬病疫苗毒株在核苷酸和推导的氨基酸水平上的同源性分别为82．0％～94．1％和87．6％-97．5％，与人用疫苗株中的CTN株G基因核苷酸和氨基酸序列同源性最高（分别为87．0％～94．1％和93．7．％～97．5％），而在兽用疫苗株中与Flury株的核苷酸和氨基酸同源性最高（分别为83．9％～84．6％和91．1％～93．0％）。在所测定序列中以来自贵州省2005年的GZ09与人用疫苗毒株CTN和动物用疫苗株Flury株G同源性最高，而以2010年的GZ30与人用疫苗毒株CTN和动物用疫苗株Flury株G基因同源性最低。系统进化树分析显示，本次所测定的序列和9株疫苗株与狂犬病病毒基因1—7型中的基因1型代表毒株聚类在同一分支，与疫苗毒株中的CTN亲缘进化关系最近，而与其他疫苗株相对较远，所测定序列中以2005年的GZ09与CTN疫苗株同源性最近，而其他序列与CTN毒株的亲缘进化关系相对较远。结论本研究从狂犬病病毒G基因水平证明贵州省狂犬病病毒流行毒株与常用的疫苗株同属狂犬病病毒基因1型，与人用疫苗株中的CTN毒株及兽用疫苗株中的Flury株差异最小，且随着时间的推移贵州省狂犬病病毒流行毒株与疫苗毒株G基因的差异越来越大，该研究结果将为狂犬病疫苗研制及有效防控措施的制定提供科学依据。
Objective To study the differences of glycoprotein gene （G gene） between rabies vi- rus epidemic strains of Guizhou province in recent years and vaccine strains, and to provide scientific basis for the development of rabies vaccine and establishment of effective control and prevention measures. Meth- ods RT-PCR assay was used to amplify the G gene of rabies positive brain tissues samples of human and dog derived from Guizhou province in recent years. The amplification products were sequenced and compara- tively analyzed with that of vaccine strains by using bioinformatics software. Results Eight full-length G gene sequences were obtained by RT-PCR amplification, sequencing and splicing. The homogeny of G gene between 8 epidemic strains of Guizhou province and 9 vaccine strains were 82.0%-94. 1% and 87.6%- 97.5% on nucleotide and deduced amino acid level, respectively, and the highest homogeny were found with the human vaccine strain CTN （87.0% -94.1% for nucleotide and 93.7. % -97.5% for amino acid） among the 6 human rabies vaccine strains, while highest homogeny were found with strain Flury （83.9% -84.6%for nucleotide and 91.1%-93.0% for amino acid ） among the three animal vaccine strains. Besides, among the 8 epidemic strains from Guizhou province, strain GZ09 collected in the year of 2005 was of the highest homogeny with human rabies vaccine strain CTN and animal rabies vaccine strain Flury, while strain GZ30 collected in the year of 2010 was of lowest homogeny with human rabies vaccine strain CTN and animal rabies vaccine strain Flury. Moreover, phylogenetic analysis based on the G gene indicated that the relationship of 8 epidemic strains derived from Guizhou, the 9 vaccine strains and genotype 1 Lyssavirus were clustered to a same branch. Vaccine strain CTN among the 9 vaccine strains was closest to the 8 epidemic strains, and the other 8 vaccine strains were relatively more distant from the epidemic strains of Guizhou province. In addi- tion, phylogenetic analysis indicated that among the 8 epidemic strains from Guizhou province, strain GZ09 collected in the year of 2005 was of closest evolutionary relationship to CTN, while the other 7 epidemic strains were relatively more distant from CTN. Conclusion This study confirmed on the G gene level that rabies virus strains circulated in Guizhou province in recent years and the vaccine strains used in China be- longed to rabies virus genotype 1, and the virus strains circulated in Guizhou province in recent years is of smallest difference with the human vaccine strain CTN and animal vaccine strain Flury. Besides, as time goes on, the difference between the epidemic strain and the vaccine strains becomes more and more obvious. The results of this study will provide scientific basis for the development of rabies vaccine and establishment of effective control and prevention measures.
出处
《中华微生物学和免疫学杂志》
CAS
CSCD
北大核心
2012年第7期595-599,共5页
Chinese Journal of Microbiology and Immunology
基金
贵州省科学技术基金项目（黔科合J字[2011]2279号）
贵州省科技厅重大专项（黔科S字[2007]1036号）
关键词
狂犬病病毒
糖蛋白基因
疫苗毒株
差异
Rabies virus
Glycoprotein gene
Vaccine strain
Differences
