应用PCR-LDR法检测乙型肝炎病毒核苷类药物耐药多基因突变位点

摘要

Objective To take the method of Multiplex polymerase chain rcaction-ligasc detection rcaction(PCR-LDR) to identify the HBV mutations on the positions of 180,181 ,184,204,236,250. Methods With the HBV DNA as templates from 50 chronic hepatitis B patients' blood scrum,PCR-LDR reaction products were obtained and detected, analyzed by ABI 3130 Sequencer. Results 8 lamivudinc-rcsistant,5 Adcfovir-resistant, 1 Entecavir-resistant and 1 lami-vudine/Adefovir-resistant cases were detected, which was consistent with the result when sequencing method was used. Conclusion PCR-LDR could be applied to fast detection of known HBV mutations,possessing advantages of easiness, convenience and celerity over other methods,thus it could help clinician to detect patients' drug resistance in the early stage and change treatments in time.%目的 用多重聚合酶连接反应-连接酶检测反应分型法(PCR-LDR)同时检测乙型肝炎病毒(HBV)的180、181、184、204、236、250位点突变.方法 以50例慢性乙型肝炎患者血清中HBV 的DNA为模板,获得PCR-LDR反应探针连接产物,用ABI 3130 Sequencer进行检测分析.结果 共检测出拉米夫定耐药突变8例(16%)、阿德福韦耐药突变5例(10%),恩替卡韦耐药突变1例(2%),拉米夫定和阿德福韦同时耐药突变1例(2%),突变情况与直接测序法检测的结果完全一致.结论 用PCR-LDR法可以快速检测HBV基因的已知位点突变,与直接测序等其他方法相比具有更简单、方便、快捷的优势,有利于临床医生早期发现乙肝病人的耐药情况并及时更改治疗方案.