四川省1株猪獾传播狂犬病病毒全基因组序列测定及分析  

Sequencing and analysis of the whole genome of a strain of rabies virus transmitted by hog badger in Sichuan province

作　　者：钟红荣 蒋明凤 冯玉亮 周兴余[1] 李伟[1] Zhong Hongrong;Jiang Mingfeng;Feng Yuliang;Zhou Xingyu;Li Wei(Sichuan Provincial Center for Disease Control and Prevention,Chengdu 610041,China)

机构地区：[1]四川省疾病预防控制中心,成都610041

出　　处：《中华实验和临床病毒学杂志》2024年第3期286-293,共8页Chinese Journal of Experimental and Clinical Virology

基　　金：四川省"十四五"生命健康重大科技专项(2022ZDZX0017)。

摘　　要：目的对四川省一例由野生动物(猪獾)咬伤发病的疑似狂犬病病例的唾液标本进行狂犬病病毒的全基因组序列测定,从分子水平对病毒进行遗传变异特征分析,了解四川省野生动物狂犬病病毒的流行和变异情况。方法从疑似狂犬病病例的唾液标本中提取总病毒RNA,采用PCR的方法以特异性引物进行狂犬病病毒序列扩增,将获得的基因片段进行测序,并运用生物学软件将所得序列进行拼接从而得到狂犬病病毒株的全基因组序列。对全基因组进行遗传变异特征分析。结果经测序获得1株猪獾源狂犬病病毒(以下简称SCR23-052)全基因组核苷酸序列,经NCBI在线BLAST及与多个参考序列比对表明,SCR23-052全基因组序列的组成和结构符合弹状病毒科狂犬病毒属特征;分别与宁夏毒株(J)、重庆毒株(CQ92、02050CHI)之间各个基因区域核苷酸与氨基酸序列相似性最高,SCR23-052基因组序列差异性在氨基酸水平明显低于核苷酸水平,说明蛋白质编码基因的核苷酸变异大多属于同义突变。种系发生分析显示SCR23-052属于基因V型,其糖蛋白主要抗原位点未见明显突变,在线网址预测糖蛋白在第56和338位发生了氨基酸糖基化,SCR23-052与参考疫苗株CTN181的信号肽序列相比氨基酸差异最少。本研究病毒在核蛋白主要抗原位点与所有参考疫苗株均发生了相同的突变(332位A→T),在B细胞表位仅与参考疫苗株CTN181相比发生了379位L→V的突变,SCR23-052与基因Ⅴ型的参考毒株在核蛋白研究位点均一致。结论获得了1株野生动物猪獾源基因V型狂犬病病毒株全基因组序列,不同于四川以往报道流行的犬源狂犬病病毒基因I型毒株。SCR23-052与各参考株基因组序列相比有不同程度差异,但主要毒力特征未改变。Objective The objective of this study is to sequence whole genome of the rabies virus in the saliva specimen of a suspected rabies case with onset of rabies from a wildlife(hog badger)bite wound in Sichuan province,where the genetic variation characteristics of the virus was analyzed at the molecular level,so as to understand the prevalence and mutation of wildlife rabies virus in Sichuan province.Methods Total viral RNA was extracted from the saliva specimen of the suspected rabies case.Then,rabies virus sequences were amplified using PCR with specific primers;the gene fragments obtained were sequenced,and the sequences obtained were spliced using biological software to obtain the whole genome sequence of the rabies virus strain.The genetic variation characteristics of the whole genome were analyzed.Results The whole genome nucleotide sequence of a strain of rabies virus of hog badger origin(hereinafter referred to as SCR23-052)was obtained by sequencing,and NCBI online BLAST and comparison with several reference sequences showed that the composition and structure of the whole genome sequence of SCR23-052 conformed to the characteristics of the Lyssavirus under the Rhabdoviridae;the highest similarity in nucleotide and amino acid sequences in various gene regions was observed between SCR23-052 and the strains of Ningxia(J)and Chongqing(CQ92,02050CHI).The sequence variability of SCR23-052 genome was significantly lower at the amino acid level than that at the nucleotide level,which indicated that most of the nucleotide variants in the protein-coding genes belonged to synonymous mutations.Phylogenetic analysis showed that SCR23-052 belonged to genotype V,which did not show any obvious mutation in the major antigenic site of the glycoprotein,underwent amino acid glycosylation at positions 56 and 338 by the online site prediction,and showed the least amino acid difference compared with the signal peptide sequence of the vaccine strain CTN181.The virus in this study has an A→T mutation at position 332 in the nucleo

关 键 词：狂犬病病毒 猪獾 全基因组 序列分析 

分 类 号：R512.99[医药卫生—内科学]