CAJ | 학술논문

目的研究拉米夫定治疗中乙型肝炎病毒(HBV)逆转录酶区核苷酸序列变异、特点、含量及其与基因型和病毒载量的关系.方法普通DNA测序法检测拉米夫定治疗的117份慢乙肝患者血清HBV逆转录酶区基因序列及其基因型;其中99份用TaqMan法定量HBV DNA;64份用焦磷酸测序(Pyrosequencing)检测YMDD基序中碱基的频率.结果 HBV YMDD变异组中C型43例,B型10例,A/B混合型1例;YMDD不变异组中C型54例,B型8例,D型1例,HBV DNA含量平均对数值在变异组和不变异组分别为:6.5699和6.6165;YMDD变异与其基因型及病毒载量无统计学意义;rtL180M位点变异与rtM204I/V位点变异高度相关;并且HBV野生株与变异株均同时存在.结论结合两种测序方法可以用来研究拉米夫定治疗中HBV基因序列变异情况及对YMDD耐药株进行定量.
목적 연구랍미부정치료중을형간염병독(HBV)역전록매구핵감산서렬변이、특점、함량급기여기인형화병독재량적관계.방법 보통DNA측서법검측랍미부정치료적117빈만을간환자혈청HBV역전록매구기인서렬급기기인형;기중99빈용TaqMan법정량HBV DNA;64빈용초린산측서(Pyrosequencing)검측YMDD기서중감기적빈솔.결과 HBV YMDD변이조중C형43례,B형10례,A/B혼합형1례;YMDD불변이조중C형54례,B형8례,D형1례,HBV DNA함량평균대수치재변이조화불변이조분별위:6.5699화6.6165;YMDD변이여기기인형급병독재량무통계학의의;rtL180M위점변이여rtM204I/V위점변이고도상관;병차HBV야생주여변이주균동시존재.결론 결합량충측서방법 가이용래연구랍미부정치료중HBV기인서렬변이정황급대YMDD내약주진행정량.
Objective To investigate the variants and quasispecies of reverse transcriptase region in polymerase gene of hepatitis B virus (HBV) during lamivudine treatment and their relationship with genotypes and viral loads. Methods HBV DNA of 117 chronic hepatitis B patients treated with lamivudine were amplified by using PCR. The PCR products including the YMDD motif were sequenced by DNA sequencer, of which, HBV DNA viral loads of 99 patients were determined by real-time PCR and 64 samples were sequenced by Pyrosequencing. Results In HBV YMDD variant group and no variant group, the HBV genotypes were 79.6% and 86. 7% of type C, 18.5% and 12. 7% of type B, 1.9% of A/B recombinant type and 2.6% of type D, respectively. The viral loads ( log 10) were 6. 5699 and 6. 6165, respectively.There was no significant difference in HBV genotypes and viral loads between these two groups. The rtL180M variant was found in association with the rtM204I/V variant, HBV variants and wild-type in YMDD motif all existed together in these two groups. Conclusions HBV variants (quasispecies) in YMDD motif could be quantified by pyrosequencing, which would be a feasible measure during nucleoside or nucleotide analogue therapy against chronic HBV infection.