成都地区HBV基因型分布研究

A study on genotype prevalence of hepatitis B virus in Chengdu area

目的调查成都地区乙型肝炎患者外周血中乙型肝炎病毒(HBV)基因型的分布情况。方法以长片段高保真聚合酶链反应(LA-PCR)扩增乙肝病毒S基因区,结合双脱氧链末端终止法(Sanger)测序技术对该地区90例乙型肝炎患者感染的HBV进行基因分型;同时与常用的基因型特异引物扩增分型法获得的分型结果进行比较,做重复试验以证实所用方法的可靠性和准确性。结果S基因直接测序法检测成都地区人群HBV基因分型结果为B基因型57例(63.3%)、C型30例(33.3%),D型3例(3.3%),无其他基因型和混合型;基因型特异引物分型法除榆出23例(25.5%)混合基因型外,其他各例结果均与S基因测序法吻合。结论成都地区HBV优势基因型以B型为主,C型次之,偶见D型。S基因直接测序法能准确鉴定HBV基因型,并较好地反映HBV基因组变化的准种特征,总体上优于型特异引物分型法,此结论具有统计学意义(P<0.001)。

Objective To investigate the prevalence of hepatitis B virus （HBV） genotypes in patients with chronic HBV infection in Chengdu area of China in combination of two different assays. Methods The HBV S gene fragment was amplified by long and accurate PCR （LA-PCR） in 90 serum samples of positive patients and then sequenced for HBV genotyping. HBV genotyping was also studied with typesFecific-primered PCR. The results from the two methods were compared to certify their credibility and accuracy. Results Genotype B was detected in 63.3%（57/90） of subjects by sequencing S fragment, genotype C in 33.3%（30/90）, genotype D in 3.3%（3/90）. None of any other genotypes in this group of patients was detected. However, mixed-genotype infection was detected in 25.5% （23/90） of subjects by genotyping following PCR with typespecific primers. Conclusion HBV genotype B was the predominant genotype in Chengdu area while genotype C was secondary, and genotype D was singularly. S fragment sequencing was believed better in genotyping HBV, which described diversity of HBV quasispecies.