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目的研究轮状病毒P[8]G1株(Wa株)在人胚肺二倍体细胞KMB17上的适应性及其免疫原性。方法将人轮状病毒P[8]G1株(Wa株)在KMB17细胞上进行适应培养,连续传代10代,免疫荧光法检测病毒的增殖情况,免疫胶体金法检测病毒的抗原性,微量滴定法检测病毒的感染性滴度,并进行病毒基因组稳定性及免疫原性检测。结果轮状病毒Wa株在KMB17细胞上连续传代后,细胞病变逐渐增快;抗原性逐渐增强;至第10代达增殖高峰,病毒滴度达4.25CCID50/ml;在传代过程中,病毒基因组核酸带型保持一致;经皮下和口服2种途径免疫小鼠,血清抗体效价均达1:8192。结论轮状病毒Wa株可在KMB17细胞上稳定增殖,病毒保持了毒株的基本生物学特性,且具有较好的免疫原性。
Objective To study the adaptability and immunogenicity of rotavirus P [8] G1 strain (Wa strain) on human embryo lung diploid cell KMB17. Methods The human rotavirus P [8] G1 strain (Wa strain) was adapted to culture on KMB17 cells for 10 generations in succession. The proliferation of the virus was detected by immunofluorescence method. The antigenicity and microtiter Method to detect the infectious titer of the virus, and the stability of the virus genome and immunogenicity testing. Results After passaged on KMB17 cells continuously, the cytopathic changes of rotavirus Wa strain gradually increased and the antigenicity gradually increased. At the tenth passage, the virus reached the peak of proliferation with a titer of 4.25 CCID50 / ml. During passage, the viral genomic nucleic acid Bands consistent; subcutaneously and orally 2 ways to immunize mice, serum antibody titers reached 1: 8192. Conclusion The rotavirus Wa strain can stably proliferate on KMB17 cells. The virus maintains the basic biological characteristics of the strain and has good immunogenicity.