Gene sequence analysis of two isolates of rabies virus epidemic in Taizhou city, Zhejiang province
摘要: 目的 分析浙江省台州市2株狂犬病病毒核蛋白及糖蛋白的基因序列,了解狂犬病病毒野毒株与人用及兽用狂犬病疫苗株间的差异。 方法 以免疫荧光法检测2008年采自台州市的144只犬脑标本,以RT-(nested)PCR法扩增病毒核蛋白及糖蛋白全基因,克隆测序后以生物信息学软件进行遗传特征分析。 结果 检测到2株狂犬病病毒阳性样品,序列分析表明两样品所携病毒均为基因1型狂犬病病毒,所携病毒的N基因核苷酸及氨基酸同源性均为100％,G基因核苷酸和氨基酸同源性分别为99.8％和99.4％。所携病毒与CTN疫苗株核苷酸同源性较高,N基因与G基因分别为88.8％和85.9％~86.1％,两样品所携病毒与Hep-Flury疫苗株的核蛋白氨基酸同源性最高,为97.6%,CTN次之,为97.1%,与CTN糖蛋白氨基酸同源性较高,为92.3%~92.5%。与诸基因1型狂犬病病毒参考株相比,两样品所携病毒与浙江温州Wz0(H)、衢州株ZJ-QZ及印度尼西亚株病毒同源性最高。系统发育分析表明XY20及JJ22与浙江温州Wz0(H)、衢州株ZJ-QZ以及印度尼西亚株、疫苗株CTN,以及泰国与马来西亚株进化关系最近,而与疫苗株aG、PV、ERA,及标准攻击毒CVS株等进化关系较远。 结论 两份阳性犬脑所携狂犬病病毒是基因1型狂犬病病毒,但无论是N基因还是G基因的核苷酸序列,以及推导出来的氨基酸序列与已知的1 型狂犬病病毒株及疫苗株存在差异。Abstract: Objective The nucleoprotein and glycoprotein gene sequences of two rabies virus strains in Taizhou city, Zhejiang province were analyzed to assess the differences between wild strains and the ones used for human and animal rabies vaccines, providing evidence for effective control of rabies. Methods Immunofluorescence assays were performed to analyze 144 dog brain samples collected from Taizhou in 2008. RT-nested PCR was carried out to amplify the entire nucleoprotein and glycoprotein genes of the virus strains. Upon the completion of cloning and sequencing, bioinformatics software was used for analysis of genetic characteristics. Results Two rabies virus strains were isolated from the samples. Sequence analysis showed that both were genotype 1 rabies viruses with 100% homology between their N gene nucleotides and amino acids, and 99.8% and 99.4% between the G gene nucleotides and amino acids, respectively. The isolates held high homologies to CTN vaccine strains in terms of nucleotides: 88.8% for the N genes and between 85.9% and 86.1% for the G genes. With respect to nucleoprotein amino acids, the two demonstrated the highest homology to Hep-Flury vaccine strains, 97.6%, followed by CTN strains, 97.1%. Their homology to CTN glycoprotein amino acids was also high (92.3%-92.5%). Among various rabies virus genotype 1 reference strains, the isolates were most homologous to Zhejiang Wenzhou Wz0(H), Quzhou ZJ-QZ and Indonesian strains. Phylogenetic analysis showed that XY20 and JJ22 were in the closest evolutionary relationship with Zhejiang Wenzhou Wz0 (H), Quzhou ZJ-QZ, Indonesia strains, CTN vaccine strains and Thailand and Malaysia strains, in contrast to a distant one with the vaccine strains, aG, PV and ERA, as well as the standard toxic attacking CVS strains. Conclusion The two rabies viruses isolated from dog brain samples belonged to genotype 1 strains, though both manifested differences from the known genotype 1 and vaccine strains in N and G gene nucleotide sequences, as well as the deduced amino acid sequences.
. Meng SL, Yan JX, Xu GL, et al. Molecular epidemiological study targeting the glycoprotein gene of rabies virus isolates from China
[J]. Virus Res,2007,124(1-2):125-138.
. Zhang Y, Xiong C, Zou Y, et al. Molecular characterization of rabies virus isolates in China during 2004
[J]. Virus Res,2006,121(2):179-188.
. Li W, He ZS, Liu LC, et al. Epidemiological study of rabies in Taizhou during 1949 - 2004
[J]. Chinese Journal of Vector Biology and Control,2007,18(1):49-51.(in Chinese) 李未, 何志胜, 刘令初, 等. 1949 - 2004年台州市狂犬病流行病学分析
. Zhang CF, Lin HQ, Zhang HF. The nurse-keeper of 34 rabies cases
[J]. Zhejiang Journal of Preventive Medicine,2008,20(6):86-88.(in Chinese)张春斐, 林荷芹, 张红枫. 34例狂犬病的护理
. Li W. Survey and study about 80 rabies cases
[J]. Modern Preventive Medicine,2008,35(3):573-577.(in Chinese)李未. 80个狂犬病病例个案调查分析
. Goto H, Minamoto N, Ito H, et al. Mapping of epitopes and structural analysis of antigenic sites in the nucleoprotein of rabies virus
[J]. J Gen Virol,2000, 81(Pt 1):119-127.
. Zhang JM, Wang LL, Yan YS, et al. Sequence analyses of the N gene in rabies viruses from Fujian province
[J]. Chinese Journal of Zoonoses,2009,25(6):515-518.(in Chinese)张建明, 王灵岚, 严延生, 等. 福建省狂犬病病毒的N基因序列分析
. Ito M, Itou T, Sakai T, et al. Detection of rabies virus RNA isolated from several species of animals in Brazil by RT-PCR
[J]. J Vet Med Sci,2001,63(12):1309-1313.
. Sato G, Itou T, Shoji Y, et al. Genetic and phylogenetic analysis of glycoprotein of rabies virus isolated from several species in Brazil
[J]. J Vet Med Sci,2004,66(7):747-753.
. Bourhy H, Kissi B, Audry L, et al. Ecology and evolution of rabies virus in Europe
[J]. J Gen Virol,1999, 80(Pt 10):2545-2557.
. Tang Q, Lillian AO, Charles ER, et al. Sequencing and position analysis of the glycoprotein gene of four Chinese rabies viruses
[J]. Chinese Journal of Virology,2000,15(1):22-33.(in Chinese)唐青, Lillian AO, Charles ER, 等. 中国四株狂犬病病毒糖蛋白基因序列分析和位点比较
. Bourhy H, Kissi B, Tordo N. Molecular diversity of the Lyssavirus genus
. Zhang YZ, Xiong CL, Zou Y, et al. An epidemiological study on rabies in Wugang and Dongkou counties, Hunan
[J]. Chinese Journal of Epidemiology,2006,27(7):583-588.(in Chinese) 张永振, 熊成龙, 邹洋, 等. 湖南省武冈市洞口县狂犬病流行病学研究
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