摘要目的在无法成功分离风疹病毒的前提下鉴定风疹病毒阳性标本的基因型,从而全面完整地了解河北省风疹病毒基因型的特征.方法采用半巢式PCR法,将盲传三代风疹病毒核酸检测为阴性而咽拭子标本风疹病毒核酸检测为阳性的风疹疑似病例咽拭子标本进行核酸扩增、序列测定和分析.结果检测的11份风疹疑似病例咽拭子标本中有6份为2B基因型风疹病毒,5份为1E基因型风疹病毒,11份标本与相应的各基因型参考株同源性高,重要抗原位点未发生变异.结论使用半巢式PCR法能够成功地鉴定出盲传为阴性的风疹病毒基因型,丰富了风疹病毒基因库,并为更全面地了解河北省风疹病毒分子流行病学提供依据.

abstractsObjective To identify the genotypes of rubella virus (RV) that had not been isolaed by cell culture,and to understand comprehensively the molecular epidemiological features of RV in Hebei province.Methods Throat swab samples which were positive by real time fluorescence quantitative PCR were cultured with three times of passaging and were tested for nucleic acid of RV.The samples with negative results after culture were selected for amplification by the semi-nested PCR and the target fragments were sequenced.Results Among the selected eleven RV throat swab samples,six were genotype 2B and five were genotype 1E.All these genotypes were highly homologous to their correspondent reference strains.No mutations were found in important sites,for example glycosylation sites,epitopes,hemagglutination inhibition and neutralization sites.Conclusions Throat swab samples can be genotyped by semi-nested PCR.This will enrich the gene sequence bank of RV and provide basis for comprehensive understanding the molecular epidemiology of RV in Hebei province.