禽冠状病毒的分离鉴定及膜蛋白基因的克隆

Isolation and identification of avian coronavirus and clone of membrane protein

目的分离鉴定禽冠状病毒,对该病毒的M基因序列进行分析。方法采用鸡胚传代分离培养病毒,用血凝特性、鸡胚半数感染量(EID50)、鸡胚矮小化、动物回归试验等方法鉴定病毒,用RT-PCR克隆病毒的膜蛋白基因并对其测序。结果分离病毒的生物学特性符合禽冠状病毒,膜蛋白基因与标准毒株膜蛋白基因为核苷酸序列同源性为91.9%,氨基酸的同源性为92.5%。结论本研究分离到1株禽冠状病毒,命名为IBV-HN05,该毒株可能是疫苗株的变异株。

Objective To isolate and identify avian coronavirus, clone and analyze the sequence of the avian coronavirus M gene. Methods A strain avian coronavirus was isolated by chicken embryo inoculation. Biological characteristics of the virus were studied by hemagglutination activity of virus to chicken red blood, EIDs0, dwarfing embryonated chicken eggs, and virus pathogenicity to chicken embryo and healthy chickens. The M gene of virus was amplified by RT-PCR, the sequence of M gene were analyzed by DNA star. Results The biological characteristics of isolating virus accorded with avian coronavirus. The nucleotide sequence homologous with standard strain was 91.9% and the amino acid sequence homologous with standard strain was 92.5%. Conclusion A strain virus named IBV-HN05 was isolated in this study, which may be an aberrance of bacterin strains.