Lei Yong-Liang, Wang Xiao-Guang, Tao Xiao-Yan, Li Hao, Meng Sheng-Li, Chen Xiu-Ying, Liu Fu-Ming, Ye Bi-Feng, Tang Qing
Lishui Center for Disease Control and Prevention, Lishui 323000, China.
Bing Du Xue Bao. 2010 Jan;26(1):45-52.
Based on sequencing the full-length genomes of four Chinese Ferret-Badger and dog, we analyze the properties of rabies viruses genetic variation in molecular level, get the information about rabies viruses prevalence and variation in Zhejiang, and enrich the genome database of rabies viruses street strains isolated from China. Rabies viruses in suckling mice were isolated, overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses from Chinese Ferret-Badger, dog, sika deer, vole, used vaccine strain were determined. The four full-length genomes were sequenced completely and had the same genetic structure with the length of 11, 923 nts or 11, 925 nts including 58 nts-Leader, 1353 nts-NP, 894 nts-PP, 609 nts-MP, 1575 nts-GP, 6386 nts-LP, and 2, 5, 5 nts- intergenic regions(IGRs), 423 nts-Pseudogene-like sequence (psi), 70 nts-Trailer. The four full-length genomes were in accordance with the properties of Rhabdoviridae Lyssa virus by BLAST and multi-sequence alignment. The nucleotide and amino acid sequences among Chinese strains had the highest similarity, especially among animals of the same species. Of the four full-length genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so the nucleotide mutations happened in these four genomes were most synonymous mutations. Compared with the reference rabies viruses, the lengths of the five protein coding regions had no change, no recombination, only with a few point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the four genomes were similar to the reference vaccine or street strains. And the four strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessed the distinct district characteristics of China. Therefore, these four rabies viruses are likely to be street viruses already existing in the natural world.
通过对4株中华鼬獾和犬源狂犬病病毒的全基因组进行测序，从分子水平分析狂犬病病毒的遗传变异特性，了解浙江省狂犬病病毒的流行与变异情况，丰富我国狂犬病病毒野毒株的基因组数据库。分离乳鼠中的狂犬病病毒，通过RT-PCR扩增重叠片段并组装全基因组，分析中华鼬獾、犬、梅花鹿、田鼠、所用疫苗株的核苷酸及推导的蛋白质相似性并进行系统发育分析。4条全基因组测序完整，遗传结构相同，长度为11923nt或11925nt，包括58nt-前导序列、1353nt-核蛋白（NP）、894nt-磷蛋白（PP）、609nt-基质蛋白（MP）、1575nt-糖蛋白（GP）、6386nt-聚合酶大蛋白（LP）以及2、5、5nt-基因间隔区（IGR）、423nt-假基因样序列（psi）、70nt-尾随序列。通过BLAST和多序列比对，4条全基因组符合弹状病毒科狂犬病病毒属的特性。中国毒株间的核苷酸和氨基酸序列相似性最高，尤其是同种动物之间。4条全基因组中，氨基酸水平的相似性显著高于核苷酸水平，因此这4个基因组中的核苷酸突变大多为同义突变。与参考狂犬病病毒相比，5个蛋白编码区的长度无变化，无重组现象，仅有少数点突变。显然，这5种蛋白似乎是稳定的。4个基因组的变异位点和类型与参考疫苗株或野毒株相似。根据多序列和系统发育分析，这4株毒株为基因1型，具有中国独特的地域特征。因此，这4株狂犬病病毒很可能是自然界中已存在的野毒株。
