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作 者:卢亦路[1] 王玲[2] 张发强[3] 陈清英[3] 夏增亮[3] 胡迪[3] 夏庆杰[1]
机构地区:[1]四川大学华西医院眼疾病分子遗传研究室,成都610041 [2]四川大学华西医院生物治疗国家重点实验室 [3]四川大学华西分子遗传研究室
出 处:《中华微生物学和免疫学杂志》2007年第6期546-548,共3页Chinese Journal of Microbiology and Immunology
摘 要: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.
关 键 词:乙型肝炎病毒 基因型 长片段高保真聚合酶链反应 S基因直接测序法 型特异 引物分型法
分 类 号:R373[医药卫生—病原生物学]
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