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目的对2014-2015年银川市流行的风疹病毒进行分离鉴定,确定其基因型,并进行基因与氨基酸的特征分析。方法采用Real-time PCR法对采集的疑似风疹病例的咽拭子标本进行初筛,阳性标本使用Vero/SLAM细胞进行病毒分离,采用RT-PCR法对病毒培养物进行风疹病毒株E1基因的标准靶核苷酸序列扩增,并对扩增产物进行序列测定与分析。结果 2014-2015年银川市共分离到24株风疹病毒株,进化树结果显示均为2B基因型;24株分离株之间的核苷酸与氨基酸同源性分别为98.1%~100.0%和97.5%~100.0%。对核苷酸编码的氨基酸序列进行比对,尽管出现了多个氨基酸的突变位点,但关键位点如N-型糖基化位点、血凝抑制位点、中和位点以及重要的抗原表位均未发生变化。结论 2014-2015年银川市流行的风疹病毒主要是2B基因型,与国内其他地区普遍流行的基因型(1E基因型)不一致。虽然出现了氨基酸的变异,但关键位点的氨基酸序列高度保守,病毒抗原性较稳定。
Objective To isolate and identify the popular rubella virus in Yinchuan City from 2014 to 2015, determine its genotypes and characterize the genes and amino acids. Methods Real-time PCR was used to screen throat swabs of suspected cases of rubella. Vero / SLAM cells were used in the positive samples for virus isolation. RT-PCR was used to determine the standard of the rubella virus strain E1 gene The target nucleotide sequence is amplified, and the amplification product is sequenced and analyzed. Results A total of 24 strains of rubella were isolated from Yinchuan in 2014-2015. The phylogenetic tree showed 2B genotypes. The nucleotide and amino acid identities of the 24 isolates were 98.1% -100.0% and 97.5%, respectively % ~ 100.0%. Alignment of nucleotide-encoded amino acid sequences, although the emergence of multiple amino acid mutation sites, but key sites such as N-type glycosylation sites, hemagglutination inhibition sites, neutralization sites and important Antigen epitopes have not changed. Conclusion The prevalence of rubella virus in Yinchuan City in 2014-2015 is mainly 2B genotype, which is inconsistent with the genotype (1E genotype) prevailing in other parts of China. Although amino acid variation occurred, the amino acid sequences of the key sites were highly conserved and the virus antigenicity was stable.