摘要
目的:基于多重PCR和反向线性杂交原理,建立乙肝病毒(HBV)耐药突变检测的新方法,探讨HBV与药物应答的关系。方法:通过序列分析,设计生物素标记的引物,扩增HBV聚合酶逆转录区基因。同时设计能检测出拉米夫定(LAM)常见耐药突变位点的特异性探针。样本经PCR扩增后与固定至尼龙膜条上的探针杂交,用BCIP/NBT系统显色,观察结果。结果:实验建立的反向线性杂交法显色后斑点出现的耐药位点与测序结果相一致。结论:检测结果与测序结果具备良好一致性,用反向线性杂交法建立的测定HBV耐药位点的具有操作简便,快速、敏感、高通量的特点。
Objective： Based on the principle of multiple PCR and reverse linear hybrid,to set up new methods of HBV drug resistance mutation detection and to discuss the relationship between HBV and drug response. Methods： By sequence analysis, biotin labeling primer was the designed, HBV polymerase gene retroviral area was amplified. Probe that can detect lamivudine （LAM） common drug resistance mutations was designed. After samples amplification by PCR and fixation to the nylon membrane probe hybridization, System BCIP/NBT coloration was applied to observe the results. Results.. The established reverse of linear hybrid method after color spots showed resistance loci consistent with sequencing results. Conclusion： Test results are consistent with sequencing results, and the reverse linear hybrid method is simple and sensitive with high flux.
出处
《海南医学院学报》
CAS
2014年第11期1482-1484,共3页
Journal of Hainan Medical University
基金
深圳市科工贸信委立项项目(201103360)
Financially Supported by Special Foundation for Clinical Studies from Committee of Medical Journal of Chinese Universities(11221022)
关键词
乙肝
反向线性杂交法
拉米夫定
耐药性
Hepatitis B Virus
Reverse linear hybrid method
Lamivudine tablets
Drug resistance
