摘要
目的对一例典型草鱼出血病的病原进行分离、鉴定和全基因组序列分析。方法采用细胞培养分离病毒、PCR检测和测序分析、电镜观察、间接免疫荧光(IFA)分析、全基因组序列测定及回归感染鉴定对采集的草鱼出血病样品进行病原分析。结果接种病毒的细胞没有产生CPE,但电镜观察可见大量呈典型呼肠孤病毒特点的病毒粒子,PCR检测并测序分析和IFA鉴定表明该分离株为II型GCRV;分离的病毒感染草鱼可出现典型的草鱼出血病症状,最终死亡率为96.7%,暂命名为GD20-08株;全基因组测序表明,GD20-08的基因组结构Ⅱ型GCRV分离株的相似,其核苷酸和氨基酸序列其他Ⅱ型GCRV分离株的同源性分别在94%~99%和96%~100%不等。结论从患草鱼出血病的草鱼体内分离到一株Ⅱ型GCRV的变异强毒株。
Objective The pathogen of a typical hemorrhagic disease of grass carp was isolated,identified and the whole genome sequence was analyzed.Methods Virus isolation by cell culture,PCR detection and sequencing analysis,electron microscope observation,indirect immunofluorescence(IFA)analysis,whole genome sequencing and regression infection experiment were used to analyze the pathogen of grass carp hemorrhagic disease samples.Results The cells inoculated with the virus did not produce CPE,but a large number of virions with typical reovirus characteristics were observed by electron microscopy.PCR detection,sequencing analysis and IFA identification indicated that the pathogen was GCRV genotype II.The isolated virus infected grass carp showed typical symptoms of grass carp hemorrhagic disease,with a mortality rate of 96.7%and tentatively named GD20-08.Whole-genome sequence analysis showed that GD20-08 was similar to other GCRV genotype Ⅱ isolates in terms of genome structure,nucleotide and amino acid sequence homology of GCRV genotype Ⅱ ranged from 94%to 99%and from 96%to 100%,respectively.Conclusion A virulent isolate of GCRV genotype Ⅱ was isolated from a grass carp suffering from haemorrhagic disease.
作者
曾伟伟
董寒旭
杨映
陈言峰
于辉
李华
ZENG Wei-wei;DONG Han-xu;YANG Ying;CHEN Yan-feng;YU Hui;LI Hua(School of Life Science and Engineering,Foshan University,Foshan 528255,China;Guangdong Provincial Key Laboratory of Animal Molecular Design and Precise Breeding,Foshan 528255,China;Key Laboratory of Animal Molecular Design and Precise Breeding of Guangdong Higher Education Institutes,Foshan 528255,China)
出处
《佛山科学技术学院学报（自然科学版）》
CAS
2022年第1期1-9,共9页
Journal of Foshan University(Natural Science Edition)
基金
广东省现代农业产业技术体系创新团队建设专项资金(2021KJ119)
佛山科学技术学院研究生自由探索基金项目(2020ZYTS35)。
关键词
草鱼呼肠孤病毒
草鱼出血病
分离
鉴定
全基因组
grass carp reovirus
hemorragic disease of grass carp
identification
whole genome
