乙型肝炎病毒分型和耐药突变基因检测应用研究

摘要

目的：研究楚雄地区乙型肝炎病毒(HBV)基因分布特征及基因耐药情况,有利于临床合理抗病毒治疗及治疗预后评估。方法采用荧光定量 PCR 和基因芯片方向斑点技术检测乙型肝炎患者血清中 DNA 水平、基因分型及耐药性。结果对103例乙型肝炎患者进行检测,未检测出乙型肝炎 DNA 水平18例,检测出 DNA 水平85例。对85例基因分型：C 型63例,占74.1%；B 型21例,占24.7%；B、C 混合型1例,占1.2%。检出野生型基因位点57例,占67%；检出突变型基因位点28例,占33%。基因突变位点 rt180M(17例),rt204V(12例),rt204I(6例),rt207I(3例),rt181V(3例),rt236T(3例)。拉米夫定5例敏感,23例耐药；替比夫定22例敏感,6例耐药；阿德福韦酯24例敏感,4例耐药；恩替卡韦敏11例敏感,17例可能产生耐药。结论该地区乙型肝炎分型主要以 C 型为主,基因突变位点主要以 rT180M 为主,其次是 rT204V。抗病毒治疗药物拉米夫定耐药最常见,替比夫定及阿德福韦酯最敏感,恩替卡韦居中。%Objective To investigate the distribution characteristics of hepatitis B virusgene types and the situation of drug re-sistance gene in Chuxiong area and to benefit the rational clinical antiviral therapy and the prognostic evaluation.Methods The DNA content,hepatitis B virus gene types and drug resistance associated sites in serum of hepatitis B patients were detected by the technology of fluorescence quantitative PCR and the gene chip of reverse dot blot.Results Among 103 serum samples,detection of DNA content in 85 samples was positive while DNA content of 18 samples was negative.85 positive samples were typed for hepati-tis B virus DNA,of which 63 samples were classified as type C(74.1%),21 samples were classified as type B(24.7%)and 1 sample was classified as mixed type B and C (1.2%).57 samples had the wild-type gene loci(67%)and 28 samples had the mutant gene locus(33%).Among these samples,1 7 cases of mutation were placed in rt180M,12 cases were placed in rt204V,6 cases were placed in rt204I,3 cases were placed in rt207I,3 cases were placed in rt181V and 3 cases were placed in rt236T.Among 28 samples of drug resistance gene mutation,5,22,24 and 1 1 samples were sensitive to lamivudine,telbivudine,adefovir and Entecavi,respectively.Con-clusion The main hepatitis B virus genotype is type C in Chuxiong area.The most important mutant site is rt180M and the second is rt204V.The most sensitive drugs are telbivudine and adefovir in clinical antiviral therapy and the second sensitive drug is Ente-cavi While the most commonly resistant drug is lamivudine.