慢性乙型肝炎病毒耐药突变体研究

在治疗慢性乙型肝炎的核苷类似物的用药过程中,筛选耐药突变体.选定1名从未接受抗病毒治疗慢性乙型肝炎患者,用抗病毒核酸类药物治疗,不同治疗时期提取血清HBV DNA,用引物P1(5′-AAGGG-TATCTTGCCCGTTTGTCGTA-3′)和P2(5′-AAGCAGGATAGCCACAGA-3′)为第1轮,P3(5′-AAGGCACTTGTAT-TCCCATCCGAG-3′)和P4(5′-AAGGTCTATTTACAGGGGA-3′)为第2轮引物扩增筛选耐药突变体.在18周和22周分别检测到突变体LMV rtH69T(YMDDlocus)和LMV rtT184T(YMDDlocus)、LMV rtM204I(YMDDlocus).在60周和70周分别检测到ADV T213S、ADV T222A、ADV K212T和ADV S196L、ADV S242H,其中ADV S196L和ADV S242H 2种突变体是首次检测到.HBV核苷酸类似物耐药突变体筛选,对研究HBV耐药的分子机制有帮助.

Drug Resistant Mutant of Chronic Hepatitis B Virus

During the remedy of chronic hepatitis B using nucleoside similitude, drug resistant mutants of HBV were screened. A chronic hepatitis patient who has never been remedied with antiviral drugs was selected and treated with antiviral nucleic acid drugs （ lamivudine and adefovir dipivoxil）, then extracted his sera HBV DNA at different reme- dy times. The drug resistant mutants were screened with amplification using P1 （5 ＇-AAGGGTATCTTGCCCGTTT- GTCGTA-3＇） P2 （5＇-AAGCAGGATAGCCACAGA-3＇） as the first turn primers, and P3 （5＇-AAGGCACTTGTATTC CCATCCGAG-3＇） P4 （5＇-AAGGTCTATTTACAGGGGA-3＇） as the second turn primers. The results showed that at 18 weeks and 22 weeks mutants of LMV rtH69T （YMDDlocus）, LMV rtT184T （YMDDlocus）, and LMV rtM204I （YMDDlocus） were detected respectively. At 60 and 70 weeks ADV T213S, ADV T222A, ADV K212, and ADV S196L, ADV S242H were detected respectively. And among them the ADV S196L and ADV S242H two mutants were discovered for the first time. Therefore, the screening of anti HBV nucleoside similitude drug resistant mutants is helpful for making clear the molecular mechanism of the HBV drug resistance.