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目的观察不同H5N1亚型禽流感病毒株空斑特性,对比研究不同空斑类型H5N1病毒致病性和复制能力差异。方法将不同稀释度的H5N1亚型禽流感病毒株分别接种于MDCK单层细胞,加盖营养琼脂,合适时间染色,观察不同H5N1亚型禽流感病毒株空斑形成特点,按照DullbeccoR的方法计算病毒PFU数;根据空斑大小进行挑选、纯化、筛选不同类型空斑病毒,测定这些病毒对Balb/c小鼠致病性差异和在MDCK细胞上复制差异。结果野生的人源H5N1禽流感病毒株在空斑形成上都存在不同程度的不均一性,空斑大小、形状差异明显:A/Vietnam/1194/2004(H5N1)以大圆形斑为主,夹杂少量小点状斑。A/Beijing/01/03(H5N1)大多数为中等大小空斑,并有少数针尖状斑。野生A/Vietnam/1194/2004经过分离纯化后,得到的两种空斑类型病毒在致病性和复制能力存在明显差异。结论野生的人源H5N1禽流感病毒株在空斑形成上都存在不同程度的不均一性,从野生病毒株中分离、纯化的大小斑病毒在致病性和复制能力上存在明显差异。
Objective To observe the plaque characteristics of different strains of H5N1 avian influenza virus and to compare the pathogenicity and replication ability of H5N1 virus with different plaque types. Methods Different dilutions of H5N1 subtype avian influenza virus strains were inoculated on MDCK monolayer cells, stained with nutrient agar and stained for a suitable time to observe the formation of plaque in different H5N1 subtype avian influenza virus strains. According to the method of DullbeccoR, virus PFU. According to the size of the plaque, we selected, purified and screened the different types of vacuolar virus. The pathogenicity of these viruses to Balb / c mice and the replication differences in MDCK cells were determined. Results The wild-type H5N1 strain developed different degrees of heterogeneity in plaque formation. The size and the shape of the plaques were significantly different: A / Vietnam / 1194/2004 (H5N1) Inclusion of a small amount of spot-like spots. A / Beijing / 01/03 (H5N1) Most are medium-sized plaques with a few needle-like spots. After isolation and purification of wild A / Vietnam / 1194/2004, the two plaque-type viruses obtained have obvious differences in pathogenicity and replication ability. Conclusions Wild-type H5N1 strain of avian influenza virus has different degrees of heterogeneity in the formation of plaque. The spot-and-spot virus isolated from the wild-type strain has obvious difference in pathogenicity and replication ability.