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为了确定耐药性甲型流感病毒H3N2神经氨酸酶的抗原性质较野生型甲型流感病毒是否发生改变,利用原核表达方法在大肠杆菌中大量表达并纯化野生型甲型流感病毒H3N2的红细胞凝集素(HA)和神经氨酸酶(NA);取HA和NA免疫后的小鼠血清分别对野生型和突变型甲型流感病毒H3N2进行Westernblotting、ELISA以及中和抗体检测。结果表明,野生型甲型流感病毒以及第119位氨基酸单点突变型甲型流感病毒对HA和NA免疫后的小鼠血清都有很好的免疫反应性,而第222位氨基酸单点突变和双点耐药突变型甲型流感病毒对免疫血清的免疫反应性要低很多。该研究首次确定了甲型流感病毒H3N2的野毒株与耐药突变株的神经氨酸酶的抗原性质存在很大的差异,进一步表明耐药突变株病毒除了具有抗药物活性之外也具有逃逸机体所产生的抗体攻击的能力,可为将来新一代甲型流感病毒H3N2诊断试剂和疫苗的开发等进一步的研究提供重要参考。
To determine whether the antigenic properties of the resistant influenza A virus H3N2 neuraminidase were altered compared to that of the wild type influenza A virus, the prokaryotic expression method was used to overexpress and purify the erythrocytes of the wild type influenza A virus H3N2 in Escherichia coli (HA) and neuraminidase (NA). Western blotting, ELISA and neutralizing antibody detection were performed on wild-type and mutant influenza A virus H3N2 respectively after sera were harvested from HA and NA-immunized mice. The results showed that the wild-type influenza A virus and the 119 amino acid single-point mutant influenza A virus had good immunoreactivity against the HA and NA-immunized mice sera, while the 222th amino acid single point mutation and The two-point drug-resistant mutant influenza A virus immunosorbent immune response is much lower. This study, for the first time, determined that the antigenic properties of the neuraminidase of the wild-type H3N2 influenza A virus and the drug-resistant mutant are quite different, further indicating that the drug-resistant mutant virus has escaped in addition to its anti-drug activity The ability of the body to produce antibody aggression provides an important reference for further research on the development of a new generation of influenza A H3N2 diagnostic reagents and vaccines.