文章摘要
两株Ⅱ型草鱼呼肠孤病毒的分离、鉴定及生物学特性比较
Comparison of biological characteristics on the two new isolates of genotype II grass carp reovirus
投稿时间:2019-06-21  修订日期:2019-09-06
DOI:
中文关键词: 草鱼出血病  草鱼呼肠孤病毒  基因II型  强弱毒株  生物学特性
英文关键词: hemorrhagic disease of grass carp  grass carp reovirus  genotype II  virulent and vaccine strain  biological characteristics
基金项目:广东省自然科学基金重点项目(2017B030311018);广东省省级科技计划项目 (2016B090918115);大宗淡水鱼产业技术体系(CARS-45-17);Ⅱ型草鱼呼肠孤病毒灭活疫苗规模化生产关键技术研究 (2019ZD0703).
作者单位E-mail
高彩霞 天津农学院 水产学院 caic2665@163.com 
曾伟伟 中国水产科学研究院珠江水产研究所 zww8810303@163.com 
汤亚方 中国水产科学研究院珠江水产研究所/农业农村部渔药创制重点实验室/广东省水产动物免疫技术重点实验室  
王英英 中国水产科学研究院珠江水产研究所/农业农村部渔药创制重点实验室/广东省水产动物免疫技术重点实验室  
王 庆 中国水产科学研究院珠江水产研究所/农业农村部渔药创制重点实验室/广东省水产动物免疫技术重点实验室  
周文礼 天津农学院 水产学院  
吴杰兴 中国水产科学研究院珠江水产研究所/农业农村部渔药创制重点实验室/广东省水产动物免疫技术重点实验室  
刘世旭 天津农学院 水产学院  
尹纪元 中国水产科学研究院珠江水产研究所/农业农村部渔药创制重点实验室/广东省水产动物免疫技术重点实验室  
李莹莹 中国水产科学研究院珠江水产研究所/农业农村部渔药创制重点实验室/广东省水产动物免疫技术重点实验室  
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中文摘要:
      为研究Ⅱ型草鱼呼肠孤病毒(GCRV)不同分离株毒力强弱及生物学特性的差异,该研究从患病和未发病草鱼体内分离到Ⅱ型GCRV各一株,分别命名为ZH1808和CQ1807,并从细胞培养特性、致病性、基因组带型、基因序列差异、遗传进化关系等方面进行比较分析。结果显示,ZH1808对草鱼和稀有鮈鲫的致死率分别为80%和100%,CQ1807对草鱼和稀有鮈鲫的致死率分别为0和10%,初步表明ZH1808为强毒株,CQ1807为弱毒株,且弱毒株感染过的草鱼对强毒株的攻击感染具有较好的免疫保护;2株分离株在草鱼上皮细胞(GSB)、稀有鮈鲫卵细胞(GRE)及稀有鮈鲫尾鳍细胞(GRF)中均能增殖但不产生致细胞病变效应(CPE),且ZH1808株的增殖量是CQ1807株的1 000倍以上;SDS-PAGE中2株分离株基因组带型相似,但S7,S8,S9,S10与S11节段的分布存在一定的差异;2株病毒的S7与S11节段的基因序列同源性分别为98%和99%。基于S7与S11节段编码的氨基酸序列构建的遗传进化树显示,2株分离株聚在同一个分支上,具有较近的亲缘关系。以上结果表明,从患病和未发病的草鱼中分离的2株II型GCRV有较多共性,但其复制能力、致病性等方面也存在较大差异。
英文摘要:
      In order to study the virulence and biological characteristics of different isolates of grass carp reovirus (GCRV) type II, we researched two new isolates of genotype II GCRV, ZH1808 and CQ1807, which were isolated from diseased and healthy grass carp respectively. The characteristics of cell culture, pathogenicity, genomic segment, gene sequence and phylogenetic differences of the two strains were compared and analyzed. The result showed that mortality rate of ZH1808 to grass carp and gobiocypris rarus were 80%, 100%, and those of CQ1807 were 10%,0% respectively. The data of mortality rate showed that ZH1808 is a virulent strain, and CQ1807 is an avirulent strain. Cell infection experiment showed the two isolates could proliferate in GSB, GRE and GRF cells, but did not cause cytopathic effect (CPE), and viral multiplication of ZH1808 strains was more than 1,000 times that of CQ1807 strains. The banding patterns of two isolates in SDS-PAGE were partially identical, but there were some differences between the segments of S7, S8, S9, S10 and S11. The sequence homology of the S7 and S11 genes between two viruses are 98% and 99%, respectively. Phylogenetic analysis indicated that isolate ZH1808 and CQ1807 are belong to the same branch, its means that they are very closely related.All results above indicate that two strains of GCRV type II isolated from diseased and healthy grass carp have more common characteristics, but their replication ability and pathogenicity are also different.
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