HBV多药耐药变异检测试剂的研制和初步评价  

The preparation and evaluation of a reagent for HBV multi-drug resistant variants detecting

作　　者：董瑞华[1,2] 张太松[2] 方凤银[2] 林炳生[1,2] 陈华云[2] 李明 程钢[1,2] 

机构地区：[1]中山大学药学院,广东广州510006 [2]中山大学达安基因诊断中心,广东广州510665

出　　处：《分子诊断与治疗杂志》2011年第2期78-82,共5页Journal of Molecular Diagnostics and Therapy

基　　金：国家科技重大专项(2008ZX10001-013);广州市科技局科技攻关重大项目(2007Z1-E4031)

摘　　要：目的研制一种基于反向点杂交技术检测HBV多药耐药变异的核酸检测试剂并对其临床应用进行初步评价。方法针对HBVP开放阅读框逆转录酶编码区的相对保守序列,设计引物探针,并设计内标和对照探针,制备HBV检测膜条,构建克隆,最后采用已构建好的克隆和50例临床样本进行检测并与测序做比较。结果克隆和50例临床样本均成功检出,反向点杂交检测结果与DNA测序结果符合率分别为100%和98%。结论本试剂具有低成本、准确度好等优点,在HBV感染辅助诊断和合理用药方面具有较好的应用前景。Objective To develop a reagent for HBV multi-drug resistant variants detecting which based on reverse dot blot hybridization（RDBH）. Methods The specific oligonucleotide probes or primers for reverse transcriptase region in HBV P ORF and internal control probes were designed and spotted on nylon membranes, a total of 12 specific clones were constructed. All the clones and 50 clinic samples were tested by using this method, and the detection results were compared with the sequence results. Results Both the clones and samples were detected successfully, and the concordance between RDBH and sequencing were 100% and 98%, respectively. Conclusion The present study shows that RDBH is an economical and accurate way for HBV multi-drug resistance detection, which can provid an impressive tool for clinical laboratory to guide the management of HBV infection.

关 键 词：乙型肝炎病毒(HBV) 多药耐药 突变 反向斑点杂交 不对称PCR 

分 类 号：R440[医药卫生—诊断学]