芯片检测HBV DNA聚合酶区基因突变及其临床应用

Detection of Polymorphism of Hepatitis B Virus P Gene by DNA Microarray

目的 基因芯片方法检测 HBV DNA聚合酶区基因突变 ,并初步探讨其临床应用价值。方法 PCR扩增 HBV DNA P区 nt45 5～ 796片断。与预制的基因芯片杂交 ,运用激光共聚焦荧光检测系统扫描芯片分析基因突变类型。结果 芯片检测 HBV DNA P区变异具有较好的特异性和准确性 ,初步应用显示 :慢性乙肝组 aa5 2 8/5 5 2突变率分别为 8.6%和 9.6% ,而无症状肝炎组和重症肝炎组突变率均为 0 ;拉米夫定治疗组与非拉米夫定治疗组比较 ,aa5 2 8突变率分别为 1 0 .5 %和 8.1 % ,aa5 5 2突变率分别为1 5 .8%和 8.1 %。结论 研究显示 HBV DNA聚合酶区基因多态性分布与病情轻重无关 ,而与病程慢性迁延有关 ;拉米夫定在耐药突变中发挥了重要作用 ;

Objective To detect HBV P gene polymorphism by D NA microarray.Methods The nt455～796 segment of HBV DNA P gene were hybridized with oligonucleotide probes,which were pre-stablized on t he chip by Cartesian Pixsys 7500.The microchip were scanned with General Scannin g 3000.According to the corresponding probe sequence,mutation type could be dete rmined.Results The preliminary experiment showed that the assay was sensitive,and specific.The mutation frequency of HBV P gene were 8.6% (aa528) and 9.6%(aa552) in chronic hepatitis B,and 0% (aa528/552) in asymptomati c carrier and severe hepatitis B,respectively.Aa528 mutation frequency of HBV P gene were 10.5% in the patients with lamivudine treatment and 8.1% in the patien ts without lamivudine treatment.Aa522 mutation frequency of HBV P gene were 15.8 % in the patients with lamivudine treatment and 8.1% in the patients without lam ivudine treatment.Conclusion The mutation frequency of HB V P gene (aa528/552) is relevent to chronic programming of hepatitis B.The lamiv udine treatment play a important role in the sites mutation.The microarray of la mivudine resistence related mutation may be useful for clinical treatment of the patients with hepatitis B.