| 悬浮Vero细胞快速扩增轮状病毒 |
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| 引用本文： | 张标,易山,张光明,谢天宏,李鸿钧,孙茂盛.悬浮Vero细胞快速扩增轮状病毒[J].粉末涂料与涂装,2010,23(5). |
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| 作者姓名： | 张标 易山 张光明 谢天宏 李鸿钧 孙茂盛 |
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| 作者单位： | 中国医学科学院北京协和医学院医学生物学研究所,昆明,650118 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 目的探讨在悬浮Vero细胞中快速扩增轮状病毒(RV)的方法。方法 Vero细胞经消化后,直接接种KMG1b7株RV。采用微量滴定法、PAGE和RT-PCR等方法比较悬浮扩增、静置培养及微载体发酵培养RV的感染性滴度、基因组核酸带型和型特异VP7基因序列;采用电镜观察和Westernblot鉴定悬浮扩增RV的形态及结构蛋白的抗原性。结果悬浮细胞扩增的RV接种24h后病毒感染性滴度达峰值,为6.00lgCCID50/ml;而静置培养和微载体发酵培养分别在接种后(96±24)h和(48±6)h达峰值,分别为(6.00±0.25)lgCCID50/ml和(6.50±0.25)lgCCID50/ml。悬浮扩增的RV基因组呈典型的4-2-3-2G1血清型带型,G1型特异基因VP7序列未发生突变。悬浮扩增的RV形态和结构蛋白的抗原性未发生改变。结论在悬浮Vero细胞中扩增RV可以缩短病毒制备周期,提高病毒收获量,有利于RV疫苗的研制或生产。
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| 关 键 词： | 轮状病毒 Vero细胞 悬浮 扩增 |
Rapid Amplification of Rotavirus in Suspension of Vero Cells |
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| ZHANG Biao,YI Shan,ZHANG Guang-ming,et al.Rapid Amplification of Rotavirus in Suspension of Vero Cells[J].Chinese Journal of Biologicals,2010,23(5). |
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| Authors: | ZHANG Biao YI Shan ZHANG Guang-ming |
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| Affiliation: | ZHANG Biao,YI Shan,ZHANG Guang-ming,et al(Institute of Medical Biology,Chinese Academy of Medical Science,Peking Union Medical College,Kunming 650118,China) |
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| Abstract: | Objective To investigate a method for rapid amplification of rotavirus(RV)in suspension of Vero cells.Methods RV KMG1 b7 strain was inoculated directly to Vero cells after digestion.The infectious titer,banding pattern of genomic nucleic acid and G1 type specific VP7 gene sequence of RV in Vero cells in suspension culture were tested by microtitration method,PAGE and RT-PCR respectively,and compared with those in stationary culture and in microcarrier fermentation culture.The morphology of RV amplified in s... |
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| Keywords: | Rotavirus(RV) Vero cells Suspension Amplification |
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