基于Ion Torrent的乙肝病毒耐药基因突变检测方法的建立与初步分析  

Drug resistance mutation analysis of Hepatitis B virus using the Ion Torrent system

作　　者：张秀娟[1] 杨金霞[1] 胡达明[1] 陈苏红[1] 周喆[1] 

机构地区：[1]军事医学科学院放射与辐射医学研究所,北京100850

出　　处：《中国体视学与图像分析》2017年第2期248-254,共7页Chinese Journal of Stereology and Image Analysis

基　　金：重大新药创制项目(2012Zx09301003005)

摘　　要：目的建立可用于指导临床用药的HBV耐药基因位点高通量测序检测方法。方法设计可覆盖全部耐药位点的区域的高通量测序建库引物,利用耐药位点区域存在差异的乙肝基因组质粒梯度混合模拟样品对高通量测序方法的突变率检测能力进行评价。结果建立了适用于ion torrent平台的扩增子文库构建方法,该方法可对HBV>100 IU/ml的血清进行建库,对模拟样中突变率在5%以上的位点可很好地检出(信噪比>10)。结论初步建立了高通量测序检测HBV耐药突变的检测方法,为临床动态监测乙肝病毒耐药区域变异、指导临床合理用药奠定了基础。Objective The objective of this study was to establish a next generation sequencing （NGS） method for HBV drug resistance mutation detecting based on an amplicon strategy. Methods The uni- versal NGS primers covering the drug resistance sites were designed based on the specific and conserved regions of HBV genome. The mixed plasmids encoded known anti - HBV drug resistance sequences was used as mimic samples to test the mutation detection efficiency of the NGS primers and NGS system. Results An amplicon strategy based on NGS method for HBV drug resistance mutation was developed. The serum with a HBV titer value higher than 100 IU/ml can be amplified by this method and mimic sam- ples with a mutation rate higher than 5% can be detected with a signal/noise ratio 〉 10. Conclusion A NGS method for detecting the anti - HBV drug resistance mutation was successfully established, which will help for guiding the rational use of anti - HBV drugs.

关 键 词：乙型肝炎 高通量测序 PGM 扩增子 

分 类 号：R373.21[医药卫生—病原生物学]