310例接受拉米夫定治疗的慢性乙型肝炎患者HBV RT区全长测序及结果分析  

Analysis of the reverse transcriptase domain of HBV polymerase in 310 patients with chronic hepatitis B under Lamivudine therapy

作　　者：任伟宏[1] 孙蕾[1] 赵素玲[1] 顾侦芳[1] 

机构地区：[1]河南中医学院第一附属医院检验科,郑州450000

出　　处：《国际检验医学杂志》2010年第1期34-36,共3页International Journal of Laboratory Medicine

摘　　要：目的对接受拉米夫定(LAM)治疗慢性乙型肝炎(CHB)患者HBV P基因RT区全长测序并分析结果。方法选择接受拉米夫定治疗6个月以上的慢性乙型肝炎患者310例,收集LAM治疗后患者的血清,采用直接PCR产物基因测序法对HBV RT区碱基进行测序,后对测序数据用Chromas 223软件进行分析,找出HBV P基因RT区基因的变异位点和相应的氨基酸,进一步分析HBV耐药突变模式。结果 1)310例患者出现位点变异的有135例(43.5%),变异位点24个;2)基础变异位点和频率:rtM204 v/I 111例(82.20%),rtS213T 8例(5.92%),rtC256S 4例(2.96%),3位点差异有统计学意义(P<0.05);3)变异模式和频率:rtM204I 39例(28.89%),rtM204V+rtL108M 25例(18.51%),rtM204V+rt204I+rtL180M 11例(8.15%),rtM204I+rtL180M 6例(4.44%),rtM204V+rtL180M+rtV173L/M 5例(3.70%),rtS213T 5例(3.70%),rtM204V+rtL180M+rtV207M/L/I 4例(2.96%),rtM204V 4例(2.96%)等;4)rtM204V还可伴随rtT184 I/S/M、rtA181T、rtC256S、rtS213T、rtL229V等位点变异。结论拉米夫定长期治疗可引起主要耐药位点rtM204 v/I突变,并常有rtL180M、rtV173L、rtS213T、rtC256S、rtV207M/L等一系列补偿性耐药位点伴随;单位点突变除rtM204I一种外,还存在rtS213T、rtM204V等突变模式;联合突变模式多数包含rtM204位点,但也存在rtS213T+rtL187I、rtC256S+rtF221Y等突变模式。Objective To study the nucleotide substitution sites and patterns of the reverse transcriptase（RT）domain of HBV potymerase of the patients with chronic hepatitis B（CHB） by means of LAM therapy in order to find the HBV resistance early. Methods the serum were selected from the patients with chronic hepatitis B,who have received the treatment of Lamivudine therapy more than 6 months, and P genes of HBV isolated from serum were amplified by means of one-step PCR and then sequenced. The difference of nucleotide,aminoacid and mutation patterns of RT region was classified by using Chromas223 software. Results 135 mutations in the HBV DNA polymerase gene of 310 patients was determined by using PCR and direct sequence. Mutation of 24 nucleotide substitution sites were found by sequencing,and the primary mutations and frequence are rtM204 v/I（n= 111, 82.20%） ,rtS213T（n=8,5.92G） ,rtC256S（n=4,2.96G）, and there is a significant difference between 3 primary mutations, mu- tation pattern and frequence：rtM204 I（n=39,28.89%）,rtM204V＋rtL180M（n= 25,18.51%）,rtM204V＋rtM204I-＋rtL180M（n =11,8.15M ）, rtM204I＋ rTL180M （n= 6,4.44%）, rtM204V＋ rtIA 80M ＋ rtV173L/M（n= 5,3. 70% ）, rtS213T（n = 5,3.70%）, rtM204V＋ rtL180M＋ rtV207M/L/I（n = 4,2.96 % ）, rtM204 V（n = 4,2. 96 % ） etc. rtM204V mutation can also be accompanied by rtT184 I/S/M,rtAI81T,rtC256S,rIS213T,rtL229V et al compensatory mutation. Conclusion The major mutations with LAM are rtM204V/I and usually companied by tL180M, rtV173L, rtS213T, rtC256S, rtV207M/L et al compensatory mutations after CHB patients received long term LAM treatment. Not only rtM204I but also rtS213T,rtM204V is of one nucleotide substitution site pattern. Most of the multi-nucleotide substitution site patterns contain rtM204,at the same time rtS213T＋rtL187I or rtC256S＋ rtF221Y can also be detected by means of sequencing.

关 键 词：肝炎 乙型 慢性 肝炎病毒 拉米夫定 抗药性 病毒 

分 类 号：R446.61[医药卫生—诊断学] R512.62[医药卫生—临床医学]