利用基因芯片分析拉米夫定治疗过程中HBV DNA的基因变异  

DNA Microarray for Monitoring Genetic Variability of Hepatitis B virus during Lamivudine Therapy

作　　者：高英堂[1] 陈瑞阳[1] 宋文芹[1] 陈成彬[1] 齐之丽[2] 景丽[2] 孙金英[2] 钱绍诚[2] 

机构地区：[1]南开大学生命科学学院,天津300071 [2]天津市第三中心医院,天津300170

出　　处：《中国病毒学》2003年第6期523-529,共7页Virologica Sinica

基　　金：天津市重大攻关资助项目(0131816111)

摘　　要：依据乙型肝炎病毒(Hepatitis B virus;HBV)聚合酶基因序列研制HBV基因芯片,此芯片可分析HBV的7个基因型、4种血清型和HBV聚合酶基因rtV173、rtL180、rtM204和rtV207位点的突变。利用此芯片对A、B两组共计45例拉米夫定治疗12个月的患者进行服药前和服药后3、6、9、12个月的动态检测,其中C基因型39例,且血清型均为adr:B基因型6例,其血清型均为adw。在完成全程检测的38例患者中,17例ALT升高的A组出现1例拉米夫定耐药变异株,而21例ALT正常的B组出现4例变异株,且所有变异株均为rtM204V/rtL180M,其中2例野生株和变异株共存。rtM204V变异最早在服药6个月时出现,随后出现rtL180M变异。10份PCR产物测序分析表明,芯片检测结果与测序结果基本一致,仅在rtL173位点出现1例差异。进一步分析HBV DNA变异与HBV DNA含量、ALT水平和HBeAg血清转换率的相关性,初步结果表明变异株的出现与治疗过程中的DNA反弹呈正相关,而与起始HBV DNA水平、ALT值无关联。HBV基因芯片可初步用于HBV DNA 检测,可能是临床追踪评价抗病毒治疗效果的较好方法之一。The Hepatitis B virus (HBV) oligochip was made according to the sequence of HBV polymerase gene. 7 genotypes and 4 sero-subtypes of HBV, as well as position rtV173, rtL180, rtM204, rtV207 in the reverse transcriptase(rt) domain of HBV polymerase,were detected with the chip.45 patients were divided into A and B groups according to their ALT levels. Serum samples for chip analysis were obtained at 0,3,6,9,12 months of treatment. Among 45 patients, 39 were genotype C and subtype adr, 6 were genotype B and subtype adw. Among 38 patients whom were treated continuously for 12 months, 1 lamivudine resistant mutant was discovered in 17 of A group with high ATL level, 4 variants were monitored in 21 of B group with normal ALT level. All variants were rtM204V and rtL180M, 2 of them were mixed with HBV wild type. The rtM204V mutant was found at 6 months of therapy, the rtL180M mutant was detected afterward. The results obtained by sequencing of the 10 PCR products and chip arraying were almost the same, the only different was that 1 variant at position rtV173 was not detected by the gene chip. Further analysing HBV DNA values, ALT levels and HBeAg seroconversion in relation to HBV mutants, the results showed that a more rapid occurrence of variant was associated with HBV DNA re-elevation, whereas not associated with HBV DNA values and ALT levels of pretreatment. The HBV gene chip could monitor genetic variability of HBV, it is a promising method for evaluating effects of lamivudine therapy.

关 键 词：基因芯片 拉米夫定治疗 基因变异 乙型肝炎病毒 

分 类 号：R346[医药卫生—基础医学]