HBV拉米呋啶耐药相关基因及HBV基因分型的初步研究

Pilot study on HBV lamivudine-resistance associated gene and HBV genotype

目的:利用基因测序技术,根据HBV S基因序列和HBV P基因区YMDD基因序列的特征,建立一种既可以诊断HBV基因型,又能检测拉米呋啶耐药突变的简便、可靠的方法。方法:采用特异的引物对待检标本进行PCR扩增后作基因序列检测,利用Chrom as2.23软件对测序结果进行分析有无突变产生,测序结果用软件DNASIS进行基因分型分析。结果:61例经拉米呋啶治疗的慢性乙肝患者B型11例(18%),C型50例(82%)。发生YMDD变异47例,变异检出率为77.0%。结论:该方法能同时检测HBV基因型和YMDD变异,可以通过准确的P基因序列测定区分YMDD变异的类型,并可根据需要监测其他的伴随突变,进一步进行核苷酸多态性分析,对临床治疗和病情监控均有指导意义。

Objective： To establish a simple and reliable method to detect HBV genotype and HBV Lamivudine-resistant mutance, which is based on HBV S gene and YMDD segment in HBV P gene by DNA sequencing. Methods： After PCR amplified the sample by the special primers, Sequenced the PCR product, analyzed the DNA sequcing data to find whether there was any lamivudine -resistant mutances or not in YMDD zone by Chromas2.23, and to confirm the genotype by the software DNASIS. Results： Eleven cases（ 18% ） were HBV B genotype,50 cases（82% ） were HBV C genotype in all 61 chronic HBV infections treated with lamivudine; and 47 cases （77%） found mutation in YMDD position. Conclusion： This method can detect HBV genotype and YMDD mutance together, and can distinguish all the 3 types of YMDD mutance with the exact DNA sequcing data of HBV P gene, and supervise the concomitant mutance according your needs. And do SNP analysis more. All these may provide instructional significations in clinical treatment and inspection the state of an illness.