【摘要】 目的对5个代次的风疹病毒疫苗株RA27/3进行全基因组序列测定与分析,以阐明该疫苗株在传代过程中基因组的稳定性。方法提取疫苗株基因组RNA,反转录合成cDNA,分为10个片段进行RT-PCR,并测定全长cDNA序列。比较这5个代次毒种之间的全基因组序列的差异。结果仅有中间代次P30和P31毒种的个别位点发生了突变,并在之后的代次中出现了恢复。P30的2429位点核苷酸的变异导致了氨基酸水平的变化,但该变化出现在基因组的非结构蛋白编码区域;而P31的3181位点核苷酸的变异未导致氨基酸水平的变化,为同义突变。结论 RA27/3疫苗株在传代过程中基因组保持了较高的稳定性,在分子水平上保证了该毒株及其生产疫苗的安全性。更多还原

【Abstract】 Objective To sequence the whole genomes of rubella virus vaccine RA27 / 3 strain of five passages and investigate their stabilities during subculture.Methods Genomic RNA of RA27 / 3 strain was extracted and reversely transcribed into cDNA,of which ten fragments were amplified by RT-PCR,and the full-length was sequenced.Whole genome sequences of the strain of five passages were compared.Results Mutations were observed only in individual sites of RA27 / 3 strain of P30 and P31,which were recovered in the passages afterwards.The mutation of nucleotide at site 2429 of P30 resulted in the change at amino acid level in the encoding region of non-structural protein of genome.However,the mutation of nucleotide at site 3181 of P31 resulted in no change at amino acid level,indicating a samesense mutation.Conclusion The genome of RA27 / 3 strain was highly stable during subculture,which ensured the safety of the strain and the prepared vaccine at molecular level.更多还原