鸭坦布苏病毒GZSS2022株分离鉴定及其全基因序列遗传进化分析  

Isolation,Identification and Genetic Evolution of the GzSS2022 Strain of the Duck Tembusu Virus

作　　者：安而立 嵇辛勤[1,2] 阮涌 陈泽辉[1,2,3,4] 曹伟 王晗晗 罗晓宇[1,2,3,4] 龙丹丹 陈佳琪 杨纯培 吴宗豪[5] 王丽娟 姚碧琼[5] AN Erli;JI Xinqin;RUAN Yong;CHEN Zhui;CAO Wei;WANG Hanhan;LUO Xiaoyul;LONG Dandan;CHEN Jiaoqi;YANG Chunpei;WU Zonghao;Wang Lijuan;YAO Biqiong(Faculty of Animal Sciences,GuiZhou University,Guiyang 550025,China;Guizhou Institute of Animal Disease,GuiZhou University,Guiyang 550025,China;Key Laboratory of Plateau Mountain Animal Genetics,Breeding and Reproduction,Ministry of Education,Guizhou University,Guiyang 550025,China;Key Laboratory of Animal Genetics,Breeding and Reproduction,Guiyang 550025,China;County Animal Disease Prevention and Control Center,Sansui 556500,China)

机构地区：[1]贵州大学动物科学学院,贵阳550025 [2]贵州省动物疫病研究所,贵阳550025 [3]贵州大学高原山地动物遗传育种与繁殖教育部重点实验室,贵阳550025 [4]贵州省动物遗传育种与繁殖重点实验室,贵阳550025 [5]三穗县动物疫病预防控制中心,三穗556500

出　　处：《病毒学报》2024年第2期392-401,共10页Chinese Journal of Virology

基　　金：贵州省科技支撑计划项目(项目号:黔科技合支撑[2020]IY040),题目:贵州三穗鸭坦布苏病毒分离鉴定及中药控制关键技术研究;贵州省科技支撑计划项目(项目号:黔科合支撑[2022]一般137),题目:三穗鸭原种场三种常发疫病净化关键技术研究与应用。

摘　　要：本研究从贵州省黔东南地区某麻鸭养殖场采集到疑似感染鸭坦布苏病毒(Duck tembusu virus,DTMUV)病的组织病料,无菌处理后接种于鸡胚连续传代培养,分离到一株病毒,通过RT-PCR、PCR鉴定及透射电镜形态观察确定该分离病毒为DTMUV,命名为GZSS2022株。为了解该分离病毒的遗传变异特点,对该分离病毒进行动物回归实验、PCR扩增及测序其全基因组、在线BLAST比对其全基因组核苷酸同源性并绘制系统发育进化树后再进行遗传进化分析。结果显示:用该病毒感染SPF雏鸭后临床症状与病理剖检基本符合DTMUV感染特征,且在病理切片中可观察到鸭脑部血管渗出淋巴细胞和单核细胞聚集在血管周围形成“血管袖套”这一典型的DTMUV感染现象。GZSS2022株与NCBI上已上传的其他DTMUV参考毒株全基因组核苷酸同源性均在96%以上,其中与广西分离株GXQZ01-2020同源性最高为99.5%,且同属于中国谱系I群毒株,与多数分离于北方地区的中国谱系II群相比,具有明显流行地域性。本次所分离的DTMUV及其全基因组序列分析结果以期为坦布苏病毒地域性流行情况与该病毒遗传变异情况的研究提供参考。Tissue samples suspected to be infected with the duck Tembusu virus(DTMUV)were collected from a duck-breeding farm in southeast Guizhou Province(China).After sterile treatment,the DTMUV was inoculated in chicken embryos for continuous culture,and a virus strain was isolated.The isolated virus was identified as DTMUV by reverse transcription-polymerase chain reaction(RT-PCR),PCR and morphological observation by transmission electron microscopy and named as the GZSS2022 strain.To explore genetic variation of the isolated virus,animal experiments were undertaken on the isolated virus.PCR amplification and sequencing of its whole genome,online BLAST comparison of the nucleotide homology of its whole genome,and analyses of phylogenetic evolution were done.Clinical symptoms and pathologic examination were basically consistent with the characteristics of DTMUV infection after infecting specific pathogen-free ducks with this virus.A typical phenomenon of DTMUV infection,"vascular cuff",was observed in pathologic sections of duck brain;blood vessels oozed lymphocytes and monocytes gathered around blood vessels.The nucleotide homology between the GZSS2022 strain and DTMUV reference strains uploaded on the National Center for Biotechnology Information database was>96%,among which homology with the Guangxi isolate GXQZ01-2020 was the highest(99.5%).This strain belonged to Chinese lineage group Ⅰ,whereas most Chinese lineage group-Ⅱ isolates were from northern China.Obvious regional prevalence was documented.Our isolation of the DTMUV and determination of its whole genome sequence could provide a reference for study of the regional prevalence and genetic variation of the DTMUV.

关 键 词：鸭坦布苏病毒 分离鉴定 全基因 遗传进化 

分 类 号：S855.3[农业科学—临床兽医学]