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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。
Objective To investigate the genome-wide sequence of five generations of rubella virus vaccine strain RA27 / 3 to clarify the genome stability of the vaccine strain during passage. Methods Genomic RNA of the vaccine strain was extracted and cDNA was reverse transcribed into 10 fragments for RT-PCR. The full-length cDNA sequence was determined. Differences in the genome-wide sequences between these 5 generations were compared. As a result, only the individual loci of P30 and P31 were mutated in intermediate generations and recovered in subsequent generations. Variation at nucleotide position 2429 of P30 resulted in a change of amino acid level, but this change occurred in the non-structural protein coding region of the genome. However, the mutation at nucleotide position 3181 of P31 did not result in the change of amino acid level Sense mutation. Conclusion The genome of RA27 / 3 vaccine maintained high stability during passage and ensured the safety of the strain and its vaccine at the molecular level.