型特异性引物PCR技术在HBV基因分型中的应用

目的 应用乙型肝炎病毒（HBV）型特异性引物PCR的基因分型方法，了解武汉地区HBV基因型的分布及其临床意义。方法 选择298例HBV感染者，分成表面抗原携带者（ASC）、急性肝炎（AH）、慢性肝炎（CH）、重型肝炎（SH）、肝硬化（LC）和原发性肝癌（HCC）6组，用多对型特异性引物PCR方法检测HBV的基因型；并对4份血清HBVDNA进行序列分析。结果多对型特异性引物PCR法可简便、快速、准确地鉴定HBV基因型；298份HBVDNA阳性的血清标本中，B基因型197例（66．1％），BC混合型68例（22．8%），C基因型33例（11．1％）；B基因型在ASC、AH、CH、SH和HCC组患者中均占绝对优势，分别为63．6％、71．9％、68．7％、89．5％和65．9%，C基因型在各组患者中的分布差异无显著性意义，BC混合基因型在各组患者中的分布存在差异，更多见于肝硬化患者；各基因型在ALT水平、HBeAg阳性率及HBV DNA水平上差异无显著性意义（均P〉0．05）。结论 多对型特异性引物PCR扩增法能快速准确地进行HBV基因分型检测，可用于大规模的流行病学调查；武汉地区HBV感染者中，优势基因型为B型，其次为BC混合型和c型，BC混合型更多见于肝硬化患者。

Study on Genotypes of HBV by PCR Using Type-specific Primers

Objective To study the distribution of hepatitis B virus(HBV) genotypes in Wuhan detected by using type-specific primers PCR and its clinical significance.Methods 298 patients with HBV infection including asymptomatic HBV carriers(ASC),acute hepatitis(AH),chronic hepatitis(CH),severe hepatitis(SH),liver cirrhosis(LC),and hepatocellular carcinoma(HCC) were collected and tested for HBV genotypes by the multiplex type-specific primers PCR.Direct sequence was performed to detect HBV DNA of 4 patients.Results A simple and precise genotyping method based on PCR using type-specific primers was developed for the determination of genotypes of HBV.Of the 298 patients,197(66.1 %) were genotype B,68(22.8 %) were mixed genotype B and C and 33(11.1 %) were genotype C.Genotype B was prevalent in ASC,AH,CH,SH and HCC patients.B and C mixed genotype in LC patients was significantly higher than others.In different genotypes the ALT value,positive rate of HBeAg and HBV DNA value had no significant difference.Conclusion Type-specific primers PCR assay is simple and precise to test HBV genotype.Genotype B is the dominant genotype of HBV,and B and C mixed genotype and genotype C exist in Wuhan.B and C mixed genotype in LC patients is significantly higher than others.There is no significant difference in the ALT value,positive rate of HBeAg and HBV DNA value among different genotypes.