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目的 建立PCR结合单酶切对戊型肝炎病毒(HEV)进行基因分型的方法,并将其应用于HEV基因型分布的研究。方法 采用简并引物扩增1～4型HEVORF1片段,1、2型HEVPCR产物为2 75、2 6 9bp ,3、4型PCR产物为317、314bp ,分别应用NaeⅠ、NotⅠ消化两种长度的PCR产物,根据酶切结果区分4种基因型。结果 采用此方法对4种基因型的HEV标准株分型,结果与预期一致;4 3份HEVIgM阳性的临床标本中,19份PCR阳性,分型结果均为4型HEV。结论 此方法可以快速简便地区分HEV 4种基因型。南京地区散发戊型肝炎大多由4型HEV所致。
Objective To establish a method for genotyping hepatitis E virus (HEV) by PCR combined with single enzyme digestion and apply it to the study of HEV genotype distribution. Methods HEVORF1 fragment of type 1 ~ 4 was amplified by degenerate primers. The PCR products of HEVPCR type 1 and 2 were 2 75 and 269 bp. The PCR products of type 3 and 4 PCR products were 317 and 314 bp. The PCR products were digested with Nae I and Not I, respectively. Products, according to the results of digestion to distinguish the four genotypes. Results The HEV standard strains of 4 genotypes were genotyped by this method, and the results were in line with the expectation. Of the 4 3 HEVIgM positive clinical samples, 19 were positive for PCR and the typing results were all of type 4 HEV. Conclusion This method can quickly and easily distinguish four genotypes of HEV. Most of hepatitis E distributed in Nanjing is caused by type 4 HEV.