一株兔出血症病毒2型鉴定与vp60基因序列分析  

Identification and Sequence Analysis of vp60 Gene of Rabbit Hemorrhagic Disease Virus Type 2

作　　者：李敏 黄云川 黄迪 李原野 周立新 石湉 叶斌 岳建国 Li Min;Huang Yunchuan;Huang Di;Li Yuanye;Zhou Lixin;Shi Tian;Ye Bin;Yue Jianguo(Chengdu Animal Disease Control Center,Chengdu 610041,China)

机构地区：[1]成都市动物疫病预防控制中心,成都610041

出　　处：《畜牧兽医科学（电子版）》2020年第14期1-4,共4页Graziery Veterinary Sciences：Electronic Version

摘　　要：兔病毒性出血症2型是由兔病毒属兔出血症病毒2型(RHDV2)引起的一种剧烈性、高度致死性兔传染病,主要病变为以实质器官出血。2010年该病在法国首次被发现,主要在欧洲流行,病兔死亡率高达80%以上。2020年4月,四川金堂县某兔场发生急性死亡病例,死亡率高达56.3%,初步怀疑为RHDV2感染。实验室对采集病料进行RT-PCR扩增、基因序列分析和病毒复制试验,结果显示其vp60基因核苷酸序列与经典RHDV的vp60基因核苷酸序列一致性为77.85%~80.94%,与RHDV2的vp60基因核苷酸序列的一致性为93.81%~97.77%,与RHDV2位于同一个进化分支。病毒复制试验显示,用病死家兔内脏组织悬液人工感染2月龄家兔(已免疫RHDV vp60株灭活苗),5只家兔在24~36 h内全部死亡,提取其内脏混合物RNA进行RT-PCR扩增并测序,结果显示其与发病兔场分离毒株一致。鉴定本次分离得到的毒株为RHDV2,并将其命名为CD202004,该毒株的发现为RHDV2检测试剂的研发和疫苗的研制提供基础。Rabbit hemorrhagic disease type 2 is a severe and highly fatal infectious disease of rabbits caused by rabbit hemorrhagic disease virus type 2(RHDV2).The main pathological changes were visceral hemorrhage.RHDV2 was first detected in France in 2010,and its rapid spread in Europe and the mortality rate is higher than 80%.In April 2020,an acute death case occurred in a rabbit farm in Jintang,Sichuan,the mortality rate was as high as 56.3%,initially suspected it was a RHDV2 infection.RTPCR,vp60 gene sequence analysis and virus replication experiment test were performed in the laboratory.The results showed that the nucleotide sequence of vp60 gene was 77.85%~80.94%identical to the vp60 gene of RHDV,and 93.81%~97.77%identical to RHDV2,which belonged to the same evolutionary branch.The virus replication test showed that five 2-month-old rabbits died within 24~36 hours after being artificially infected with the suspension of diseased rabbit viscera(vp60 strain of inactivated vaccine with RHDV).Their offal mixture was detected by RT-PCR and sequenced.The results showed that it was consistent with the virus isolated from the infected rabbit farm.In conclusion,the isolated strain was identified as RHDV2 and named as CD202004,This study provides a basis for the development of RHDV2 Detection Reagent and vaccine.

关 键 词：兔病毒性出血症2型(RHDV2) VP60基因 RT-PCR 序列分析 

分 类 号：S852.65[农业科学—基础兽医学]