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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: To establish a high-throughput sequencing method for HBV resistance gene loci that can be used to guide clinical medication. Methods High-throughput sequencing primers were designed to cover all the drug-resistant loci. The ability of mutation detection of high-throughput sequencing was evaluated by using plasmid gradient mixed analogue samples of hepatitis B genome with different drug resistance loci. Results The construction of an amplicon library suitable for ion torrent platform was established. This method can build a library of sera with HBV> 100 IU / ml, and can detect the sera with the mutation rate above 5% in the simulated samples Out (signal to noise ratio> 10). Conclusion A high-throughput sequencing method for detection of HBV resistance mutations was initially established, which laid the foundation for the clinical dynamic monitoring of HBV variability and guiding clinical rational drug use.