【摘要】 目的:研究乙肝病毒不同复制期与乙型肝炎患者抗病毒用药后基因突变两者的相关性。方法:选取203例慢性乙型肝炎患者血清标本为对象。经巢式PCR扩增及磁珠分离,制备焦磷酸测序单链模板,将PCR扩增得到HBV病毒P区和BCP区产物经焦磷酸测序进行突变频率检测,并在PyroMark ID遗传分析系统上进行焦磷酸测序。结果:检测乙型肝炎病毒DNA含量,按照HBV定量结果阳性(拷贝数≥103copies/ml)和阴性(拷贝数<103copies/ml)的标本分组分析,HBV定量阳性标本具有极高的敏感性。批量突变检测,乙型肝炎病毒复制率越高,突变率越高,检测率越高。结论:选择乙肝病毒的复制阳性标本,结合焦磷酸测序进行高通量、快速、准确检测耐药基因区突变,能满足临床批量用药检测及治疗方案设计的需要。更多还原

【Abstract】 Objective To explore the correlation between the different period of hepatitis B virus(HBV) replication and genetic mutation with hepatitis B virus after medication in patients.Methods The serum of 203 patients with chronic hepatitis B(CHB) were determined by nested PCR and were separated by magnetic bead,to prepare single-stranded template of pyrosequencing.Mutation frequency of PCR amplified HBV viral P and BCP area were detected by pyrosequencing,which used PyroMark ID genetic analysis systems.Results According to the results of hepatitis B virus DNA levels,the serum were separated HBV-positive group(copy number ≥103 copies/ml) and negative group(copy number <103 copies/ml).HBV-positive samples have a high quantitative sensitivity.With the rate increaseing of hepatitis B virus replication,the higher of mutation rate,the higher detection rate.Conclusion Selected the hepatitis B virus-positive samples of replicating.Using high-throughout pyrosequencing way may rapidly accurate fo detect resistance mutations in genes,it is helpful to satisfy demand for clinical drug testing and design of treatment programme.更多还原