河南农业科学 ›› 2025, Vol. 54 ›› Issue (11): 144-151.DOI: 10.15933/j.cnki.1004-3268.2025.11.015

• 畜牧·兽医 • 上一篇    下一篇

1 株重组鹅细小病毒的分离鉴定及其基因组序列分析

薛拥志1,2,雷白时1,3,刘曼迪1,路文彬1,赵款1,3,4,张武超1,3,胡满1,袁万哲1,3,4   

  1. (1. 河北农业大学 动物医学院,河北 保定 071000;2.河北北方学院 动物科技学院,河北 张家口 075000;3.河北省兽医生物技术创新中心,河北 保定 071000;4.国家兽用生物制品工程技术研究中心,江苏 南京 210014)
  • 收稿日期:2025-07-07 接受日期:2025-08-26 出版日期:2025-11-15 发布日期:2025-11-24
  • 通讯作者: 袁万哲(1978-),男,陕西扶风人,教授,博士,主要从事动物传染病致病机制研究。E-mail:yuanwanzhe@126.com 胡满(1962-),男,河北保定人,教授,博士,主要从事动物免疫学研究。E-mail:manhu1962@163.com
  • 作者简介:薛拥志(1977-),男,河北隆尧人,副教授,博士,主要从事病原微生物的分子生物学及致病机制研究。E-mail:xyzh772000@163.com
  • 基金资助:
    “十四五”国家重点研发计划项目(2023YFD1802602);河北省省属高等学校基本科研业务研究项目(KY2021007);河北省在读研究生创新能力培养资助项目(CXZZBS2024083);河北农业大学科研发展基金计划项目(JY2022026);石家庄市科技计划项目(226500427A)

Isolation,Identification and Genomic Characterization Analysis of a Recombinant Goose Parvovirus Strain

XUE Yongzhi1,2,LEI Baishi1,3,LIU Mandi1,LU Wenbin1,ZHAO Kuan1,3,4,ZHANG Wuchao1,3,HU Man1,YUAN Wanzhe1,3,4   

  1. (1.College of Veterinary Medicine,Hebei Agricultural University,Baoding 071000,China;2.College of Animal Science and Technology,Hebei North University,Zhangjiakou 075000,China;3.Hebei Veterinary Biotechnology Innovation Center,Baoding 071000,China;4.National Research Center of Engineering and Technology for Veterinary Biologicals,Nanjing 210014,China)
  • Received:2025-07-07 Accepted:2025-08-26 Published:2025-11-15 Online:2025-11-24

摘要: 为明确河北省定兴县鹅细小病毒(GPV)流行毒株的基因组特征及遗传进化规律,对来自河北省定兴县某鹅场35日龄疑似小鹅瘟致死鹅的肝脏病料进行病原分离与鉴定。经鹅胚尿囊腔接种并培养病毒,通过透射电镜(TEM)观察病毒粒子形态;利用PCR分段扩增并测序,经拼接获得一株GPV分离株的全基因组序列;进一步通过序列比对分析其基因组结构,预测潜在糖基化位点,并开展重组检测与系统发育分析。结果表明,该分离株在鹅胚培养中可致尿囊液内产生大量尿酸盐;透射电镜观察可见直径约20 nm的病毒颗粒。动物致病性试验结果表明,该毒株对雏鹅的致死率达53.3%。全基因组测序及分析显示,分离株基因组全长为5 046 bp,命名为HB-DX株(GenBank登录号:OR544341.1)。序列比对发现,HB-DX株与RC16、82-0321、YZ99-6等毒株类似,在基因组两端的UTR区均存在两处14 bp的缺失。糖基化位点预测结果显示,该毒株非结构蛋白1(NS1)编码区存在4个潜在糖基化位点,结构蛋白1(VP1)编码区存在5个潜在糖基化位点。重组分析表明,HB-DX株发生2次基因重组事件,其主要亲本为欧洲致病性B株,次要亲本为我国疫苗株SYG61v。遗传进化分析显示,该分离株与GPV分支毒株的核苷酸同源性为93.3%~99.6%,与NGPV(新型鹅细小病毒)分支为92.9%~94.4%,而与MDPV(番鸭细小病毒)分支的同源性较低,为80.0%~84.1%。综上,HB-DX为一株经重组产生的强毒力GPV毒株。

关键词: 鹅细小病毒, 分离鉴定, 糖基化位点, 基因重组分析

Abstract: To elucidate the genomic characteristics and genetic evolutionary patterns of the prevalent goose parvovirus(GPV) strain in Dingxing County,Hebei Province,the pathogen was isolated and identified from liver tissue samples of 35‑day‑old geese suspected of dying from gosling plague in a goose farm located in Dingxing County. The virus was inoculated into the goose embryo allantoic cavity,and the morphology of the virus particles was observed by transmission electron microscopy(TEM). The whole genome sequence of a GPV isolate was obtained by PCR amplification and sequencing. The genome structure was further analyzed by sequence alignment,potential glycosylation sites were predicted,and recombination detection and phylogenetic analysis were carried out. The results showed that this isolate caused the production of large amounts of uric acid salts in the allantoic fluid during goose embryo culture;TEM observation revealed viral particles with a diameter of approximately 20 nm.Animal pathogenicity tests indicated a 53. 3% mortality rate in goslings. Whole genome sequencing and analysis revealed that the isolate had a genome length of 5 046 bp and was named HB‑DX strain(GenBank accession number:OR544341.1). Sequence alignment revealed that the HB‑DX strain was similar to RC16,82‑0321,YZ99‑6 and other strains,and there were two 14 bp deletions in the UTR region at both ends of the genome. The prediction of glycosylation sites showed that the non‑structural protein 1(NS1)coding region of this strain contained 4 potential glycosylation sites,while the structural protein 1(VP1)coding region contained 5. Recombination analysis indicated that the HB‑DX strain underwent two recombination events,with the European pathogenic B strain as the major parent and the Chinese vaccine strain SYG61V as the minor parent. Phylogenetic analysis showed that the nucleotide homology of this isolate was 93.3%—99.6% with GPV branch,and 92.9%—94.4% with NGPV(novel goose parvovirus)branch,while its homology was significantly lower at 80.0%—84.1% with MDPV(muscovy duck parvovirus) branch. In conclusion,HB‑DX is a highly virulent GPV strain originated through recombination.

Key words: Goose parvovirus, Isolation and identification, Glycosylation site, Gene recombination analysis

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