Lei Yong-Liang, Wang Xiao-Guang, Liu Fu-Ming, Chen Xiu-Ying, Ye Bi-Feng, Mei Jian-Hua, Lan Jin-Quan, Tang Qing
Lishui Center for Disease Control and Prevention, Zhejiang 323000, China.
Zhonghua Liu Xing Bing Xue Za Zhi. 2009 Aug;30(8):824-8.
Based on sequencing the full-length genomes of two Chinese Ferret-Badger, we analyzed the properties of rabies viruses genetic variation in molecular level to get information on prevalence and variation of rabies viruses in Zhejiang, and to enrich the genome database of rabies viruses street strains isolated from Chinese wildlife.
Overlapped fragments were amplified by RT-PCR and full-length genomes were assembled to analyze the nucleotide and deduced protein similarities and phylogenetic analyses of the N genes from Chinese Ferret-Badger, sika deer, vole, dog. Vaccine strains were then determined.
The two full-length genomes were completely sequenced to find out that they had the same genetic structure with 11 923 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 two 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 two full-length genomes, the similarity in amino acid level was dramatically higher than that in nucleotide level, so that the nucleotide mutations happened in these two genomes were most probably as synonymous mutations. Compared to the referenced rabies viruses, the lengths of the five protein coding regions did not show any changes or recombination, but only with a few-point mutations. It was evident that the five proteins appeared to be stable. The variation sites and types of the two ferret badgers genomes were similar to the referenced vaccine or street strains. The two strains were genotype 1 according to the multi-sequence and phylogenetic analyses, which possessing the distinct geographyphic characteristics of China. All the evidence suggested a cue that these two ferret badgers rabies viruses were likely to be street virus that already circulating in wildlife.
通过对两只中华鼬獾的狂犬病病毒全长基因组进行测序，从分子水平分析狂犬病病毒遗传变异特性，了解浙江省狂犬病病毒的流行和变异情况，丰富我国野生动物源狂犬病病毒街毒株的基因组数据库。
采用RT-PCR扩增重叠片段，拼接全长基因组，分析中华鼬獾、梅花鹿、田鼠、犬源狂犬病病毒N基因的核苷酸及推导氨基酸相似性，并进行系统进化分析，确定疫苗株。
成功测定了两个全长基因组序列，发现它们具有相同的基因结构，全长11923nt，包括58nt的前导序列、1353nt的核蛋白（NP）基因、894nt的磷蛋白（PP）基因、609nt的基质蛋白（MP）基因、1575nt的糖蛋白（GP）基因、6386nt的聚合酶大蛋白（LP）基因，以及2、5、5nt的基因间隔区（IGR）、423nt的假基因样序列（Psi）、70nt的尾随序列。
经比对和多序列比对分析，两个全长基因组符合弹状病毒科狂犬病病毒属的特性。我国分离株之间核苷酸和氨基酸序列相似性最高，尤其是同种动物之间。两个全长基因组氨基酸水平的相似性显著高于核苷酸水平，推测这两个基因组中的核苷酸突变很可能为同义突变。与参考狂犬病病毒相比，五个蛋白编码区的长度未发生改变或重组，仅存在少量点突变，表明这五个蛋白较为稳定。两个鼬獾基因组的变异位点和类型与参考疫苗株或街毒株相似。经多序列和系统进化分析，这两个毒株属于基因1型，具有我国独特的地理特征。所有证据提示，这两株中华鼬獾狂犬病病毒很可能是已在野生动物中传播的街病毒。
