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为了解拉米夫定治疗后发生酪氨酸-蛋氨酸-天冬氨酸-天冬氨酸(Tyrosine(Y)-methionine(M)-aspartic acid(D)-aspartic acid(D),YMDD)基序变异慢性乙肝患者乙肝病毒逆转录基因(Reverse transcriptase,RT)变异特点及对与之相重叠的表面抗原基因区(Surface antigen region,S)变异的影响,为乙肝的诊断和治疗理论提供依据。本研究应用聚合酶链反应(Polymerase chain reaction,PCR)方法对25例发生YMDD变异慢性乙肝患者和30例未进行抗病毒治疗的慢性乙肝患者乙肝病毒(Hepatitis B virus,HBV)基因组RT基因进行扩增,扩增产物进行直接测序分析。结果显示发生YMDD变异患者rt204位点变异类型分布以rtM204I为主(20/25,80%),多伴有rtL80I变异(14/20,70%);rtM204V变异同时伴有rtL180M变异(5/5,100%)。YMDD变异患者RT区变异发生率高于未治疗患者(P<0.05)。YMDD变异患者在5个经典核苷类耐药相关位点(rtL80,rtV173,rtL180,rtM204和rtM250)存在变异。两组患者在其它19个潜在核苷类耐药相关位点存在变异,而YMDD变异患者RT区潜在核苷类耐药相关位点变异频率显著高于未治疗患者(P<0.05),而且还存在对其它(拉米夫定以外)核苷类药物耐药相关位点的变异。发生YMDD变异患者S区变异发生率高于未治疗患者(P<0.05),在YMDD变异患者的S区“a”决定簇中还出现了sM133L和sG145R变异。研究表明拉米夫定治疗后发生YMDD变异患者HBV基因组RT区除发生rtM204I/V变异外还发生了其它耐药相关位点变异,可能导致对其它核苷类的交叉耐药。同时RT区变异影响重叠的S基因区造成HBsAg变异,可能造成对HBsAg检测的影响和免疫逃避的发生。
In order to understand that Tyrosine (Y) -methionine (M) -aspartic acid (D) -aspartic acid (D), YMDD) group occurs after lamivudine treatment The variation of hepatitis B virus reverse transcriptase (RT) and its effect on the variation of surface antigen region (S) in sequence-variant chronic hepatitis B patients provide the basis for the diagnosis and treatment of hepatitis B. In this study, polymerase chain reaction (Polymerase chain reaction, PCR) method of 25 patients with YMDD mutation in chronic hepatitis B patients and 30 patients without chronic hepatitis B virus treatment of hepatitis B virus (Hepatitis B virus, HBV) RT gene amplification Increase, amplification products for direct sequencing analysis. The results showed that the distribution of rt204 mutation in patients with YMDD mutation was mainly rtM204I (20 / 25,80%), with rtL80I mutation (14/20 and 70%), rtM204V mutation was associated with rtL180M mutation (5 / 5,100 %). The mutation rate of RT in YMDD mutation patients was higher than that of untreated patients (P <0.05). YMDD mutations in five classical nucleotide-resistant nucleotide-related sites (rtL80, rtV173, rtL180, rtM204 and rtM250) there is variation. Mutation in the other 19 potential nucleoside resistance-related sites was found in both groups, but mutation frequency of potential nucleoside resistance-related sites in RT region of YMDD mutation patients was significantly higher than that of untreated patients (P <0.05) There are variability in sites related to resistance to other nucleosides (other than lamivudine). The incidence of S-region mutation in patients with YMDD mutation was higher than that in untreated patients (P <0.05), and sM133L and sG145R mutations were also found in the S region “a” determinant in patients with YMDD mutation. Studies have shown that patients with YMDD mutation after lamivudine treatment in addition to rtM204I / V mutations occurred in the HBV genomic RT also occurred in other resistance-related site mutations, may lead to cross-resistance to other nucleosides. At the same time, mutations in the RT region affect the overlap of the S gene region caused by HBsAg mutation, which may result in HBsAg detection and immune evasion.