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浙江省1株犬源狂犬病病毒全基因组序列测定及细胞分离鉴定北大核心CSCD
引用本文:张冉昕,马岩,赵中飞,任江萍,凌锋,卢学新,朱武洋. 浙江省1株犬源狂犬病病毒全基因组序列测定及细胞分离鉴定北大核心CSCD[J]. 中国人兽共患病杂志, 2022, 38(10): 849-853. DOI: 10.3969/j.issn.1002-2694.2022.00.130
作者姓名:张冉昕  马岩  赵中飞  任江萍  凌锋  卢学新  朱武洋
作者单位:1.包头医学院公共卫生学院,包头 014000;
2.中国疾病预防控制中心病毒病预防控制所,北京 102206;
3.绍兴市疾病预防控制中心,绍兴 312000;
4.诸暨市疾病预防控制中心,绍兴 311800;
5.浙江省疾病预防控制中心,杭州 310051
基金项目:国家重点研发计划(No.2019YFC1200701)
摘    要:目的了解浙江地区狂犬病分子流行病学现状,为浙江地区狂犬病防控提供相关数据。方法使用RT-PCR扩增浙江狂犬病病毒街毒株核苷酸并进行全基因组序列测定,与32株中国狂犬病病毒街毒株基因序列进行比对分析;使用Neuro-2a细胞进行病毒分离,测定不同代次全基因组序列并比较核苷酸突变情况。结果获得了全长为11782 bp的全基因组序列,经过比对分析后发现该毒株属于ChinaⅠ型,与全部参考基因Ⅰ型病毒核苷酸序列一致性为97.10%~99.31%;通过Neuro-2a细胞成功分离到狂犬病病毒,连续三代培养后病毒滴度最终达到5.71×10^(7)FFU/mL,子代之间序列一致性在99.9%以上,其中F3代较F1代核苷酸序列发生了7个位点突变,氨基酸未发生改变。结论新狂犬病病毒分离毒株(ZJSX-2021)属于ChinaⅠ型,与以往分离毒株属同一基因型,表明狂犬病病毒近年在浙江地区仍持续传播。细胞分离后病毒序列并无明显变化。

关 键 词:狂犬病病毒  全基因组  序列分析  细胞培养
收稿时间:2021-12-15

Identification of a canine rabies virus in Zhejiang Province through cell isolation and complete genome sequencing
ZHANG Ran-xin,MA Yan,ZHAO Zhong-fei,REN Jiang-ping,LING Feng,LU Xue-xin,ZHU Wu-yang. Identification of a canine rabies virus in Zhejiang Province through cell isolation and complete genome sequencing[J]. Chinese Journal of Zoonoses, 2022, 38(10): 849-853. DOI: 10.3969/j.issn.1002-2694.2022.00.130
Authors:ZHANG Ran-xin  MA Yan  ZHAO Zhong-fei  REN Jiang-ping  LING Feng  LU Xue-xin  ZHU Wu-yang
Affiliation:1. School of Public Health, Baotou Medical College, Baotou 014000, China;
2. National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, Beijing 102206, China;
3. Shaoxing Center for Disease Control and Prevention, Shaoxing 312000, China;
4. Zhuji Center for Disease Control and Prevention, Shaoxing 311800, China;
5. Zhejiang Center for Disease Control and Prevention, Hangzhou 310051, China
Abstract:This study aimed to understand the molecular epidemiology of rabies virus, and to provide relevant data for rabies prevention and control in Zhejiang Province. Nucleotides of Zhejiang street rabies virus strains were amplified by RT-PCR, and the whole genome sequence was determined and compared with the gene sequences of 32 Chinese street rabies virus strains. After viral isolation with Neuro-2a cells, the whole genome sequences of several generations were determined, and the nucleotide mutations were compared. A full-length genome sequence of 11 782 bp was obtained. After comparison and analysis, the strain was identified as China I, and the nucleotide sequence consistency with genotype I for all reference genes was 97.10%-99.31%. The virus strain was successfully isolated with Neuro-2a cells, and after three generations of culture, the titer reached 5.71×107 FFU/mL. The sequence identity among the progeny exceeded 99.9%. The F3 generation had seven site mutations with respect to the F1 generation in the nucleotide sequence, but the amino acid sequence did not change. The new rabies virus isolate (ZJSX-2021) belonged to China I and the same genotype as the previously isolated strains, thus indicating that rabies virus has continued to spread in Zhejiang Province in recent years. No significant change in virus sequence was observed after cell isolation, thus providing a basis for the study of rabies virus in Zhejiang Province.
Keywords:rabies virus  whole genome  sequence analysis  cell culture  
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