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本研究对2001～2011年本实验室保存的血清标本检测结果为麻疹IgM阴性,风疹IgM阳性病例相对应的咽拭子标本进行风疹病毒(Rubella virus,RV)分离,用RT-PCR方法对细胞培养产物鉴定后扩增病毒E1基因,扩增产物用于核苷酸序列测定,并进行分子流行病学分析。结果表明,60份咽拭子标本共分离到31株RV,获得27株RV的739nt(nt8731～nt9469)核苷酸序列,系统同源性分析表明,27株RV分离株分别属于两个不同的基因型,除分离自2011年的11009株、11052株和11106株为2B基因型外,其他分离株均为1E基因型。27株RV分离株大部分的核苷酸突变为无义突变,氨基酸序列高度保守。24株1E基因型分离株中大部分毒株氨基酸序列完全一致,自2001年以来可能有来自同一传播链的RV在持续传播。本研究首次监测到2B基因型RV2011年开始在上海流行,经GenBank核苷酸序列比对发现,其与越南、日本、阿根廷等国家近几年的RV分离株核苷酸序列同源性达99%。由于以前对风疹的监测较少,尚不能证明其来源。
In this study, serum samples from our laboratory were detected for Rubella virus (RV) isolates from measles-negative IgM and rubella virus (IgM) -positive cases from 2001 to 2011. RT- After culturing the product for identification, the virus E1 gene is amplified, and the amplified product is used for nucleotide sequence determination and molecular epidemiological analysis. The results showed that a total of 31 RV isolates were isolated from 60 throat swabs and 739 nt (nt8731 ~ nt9469) of 27 RVs were obtained. The phylogenetic analysis showed that 27 RV isolates belonged to two different The genotypes were all 1E genotypes, except for the isolates of genotype 2B from 11009, 11052 and 11106 strains isolated in 2011. Most of the 27 RV isolates mutated to nonsense mutations, and their amino acid sequences were highly conserved. The amino acid sequences of most strains of 24 isolates of 1E genotypes were completely consistent. RVs from the same transmission chain may have been continuously transmitted since 2001. In the present study, 2B genotype RV was first detected in 2011 in Shanghai. According to the GenBank nucleotide sequence alignment, the nucleotide sequence homology with RV isolates of Vietnam, Japan and Argentina in recent years was 99 %. Due to less previous monitoring of rubella, its source can not yet be proven.