【摘要】 目的探讨慢性乙型肝炎患者经核苷(酸)类似物抗病毒治疗后出现短暂性病毒学突破与逆转录酶区基因变异的关系。方法在随访门诊中抽取16例接受核苷(酸)类似物抗病毒治疗出现短暂性病毒学突破者的预留血清,并提取HBV DNA,采用巢式PCR方法扩增HBV逆转录酶区基因,PCR产物回收纯化后直接测序,将直接测序结果用DNASTAR进行分析,找出HBV逆转录酶区基因的变异位点和相应的氨基酸。结果 16例短暂性病毒学突破患者HBV逆转录酶区均出现不同程度的点突变,其中主要的变异位点rtI/F122L、rtQ130P、rtS135Y、rtK149Q、rtW275替换为终止密码子(TAG)、rtV266I、rtH/Q271E的发生频率分别为56.3%、37.5%、43.8%、56.3%、50.0%、37.5%和37.5%,但无一例出现既往报道的常见耐药位点(rtI169T、rtV173L、rtL180M、rtA181V/T、rtT184G、rtA194T、rtS202I/G、rtM204I/V、rtI233V、rtN236T、rtM250V/L)变异。结论短暂性病毒学突破者逆转录酶区可出现不同程度的核苷酸位点变异及氨基酸替换,但均未检测到核苷(酸)类似物相关的常见耐药位点的变异。更多还原

【Abstract】 Objective To determine the correlation between the transient virological breakthrough(tVBT) in chronic HBV patients receiving nucleos(t)ide analogues(NUCs) and the mutation of reverse transcriptase domain of HBV in clinical practice.Methods HBV DNA in serum samples of 16 HBV patients with tVBT were extracted for amplification of the reverse transcriptase region of HBV genome by nested-PCR.The mutated sites of sequenced reverse transciptase region were alighned with Seqman software(DNASTAR inc.,Madison WI).Results Mutation of 17 nucleotide substitution sites were found by sequencing,and the primary mutations(frequency) were rtI/F122L(56.3%),rtQ130P(37.5%),rtS135Y(43.8%),rtK149Q(56.3%),rtV266I(37.5%),rtH/Q271E(37.5%) and termination codon TAG replacing the rtW275(50.0%).However,there were no evidence of common NUCs drug resistance mutations were revealed(rtI169T,rtV173L,rtL180M,rtA181V/T,rtT184G,rtA194T,rtS202I/G,rtM204I/V,rtI233V,rtN236T and rtM250V/L).Conclusion In patients with trVBT during NUCs therapy,mutations of nucleotide substitution sites are found in the reverse transcriptase domain of HBV,but with no relation to antiviral drug resistance.更多还原