论文部分内容阅读
目的产生在Vero(非洲绿猴肾)细胞中能高适应生长的重配H5N1流感病毒疫苗株,并对其生物学特性进行初步测定。方法修饰A/Puerto Rico/8/34(H1N1)(PR8)的NS基因为Vero细胞适应型,并合成NS基因片段,构建质粒pHW2000-NS;通过RT-PCR方法,扩增疫苗株A/Anhui/01/2005(H5N1)的HA、NA基因片段,构建质粒pHW2000-HA、pHW2000-NA;以PR8的其它5个内部基因作为骨架,按照1+2+5模式8质粒共转染Vero细胞拯救流感病毒,并对拯救病毒的生物学特性进行检测。结果由Vero拯救出具有高适应生长特性的H5N1亚型人禽流感病毒疫苗株,并检测了重配疫苗株的生物学特性。结论成功从Vero细胞拯救了具有Vero细胞高适应性生长特性的H5N1亚型人禽流感病毒疫苗株,为应用Vero细胞大规模生产流感疫苗奠定了基础。
Objective To generate a vaccine strain of reassortant H5N1 influenza virus capable of highly adaptive growth in Vero (African green monkey kidney) cells and to determine its biological characteristics. Methods The NS gene of A / Puerto Rico / 8/34 (H1N1) (PR8) was modified to Vero cells and the NS gene fragment was synthesized to construct plasmid pHW2000-NS. The vaccine strain A / Anhui was amplified by RT-PCR HA / NA gene fragment of H5N1 / 01/2005 (H5N1) was constructed to construct plasmids pHW2000-HA and pHW2000-NA. Vero cells were co-transfected with the other 5 internal genes of PR8 Influenza virus, and to detect the biological characteristics of the rescue virus. As a result, the H5N1 subtype H5N1 virus vaccine strain with high adaptive growth characteristics was rescued by Vero and the biological characteristics of the reassorted vaccine strains were examined. Conclusions The H5N1 subtype H5N1 virus vaccine strain with Vero cell highly adaptable growth characteristics was successfully rescued from Vero cells, which laid the foundation for the large-scale production of influenza vaccine using Vero cells.