摘要
目的 探讨 ＨＢｅＡｇ阴性的慢性重型乙型肝炎患者ＨＢＶ前Ｃ区变异及前Ｃ区 Ｔ１８６２突变体对ｅ抗原合成和分泌影响。方法选取９例重型乙型病毒性肝炎患者，用ＰＣＲ方法扩增血清中ＨＢＶ前Ｃ／Ｃ基因区，并克隆后测序和序列分析；构建ＨＢＶ前Ｃ区Ｔ１８６２位点突变体表达质粒ｐＧＥＭＴ，经体外翻译比较野毒株和变异株的表达产物。结果ＨＢＶ前Ｃ区有３个位点变异使氨基酸序列改变，Ａ１８９６、 Ａ１８９９和Ｔ１８６２；Ａ１８９９并不单独出现。前Ｃ区Ｔ１８６２突变并不阻止ＨＢＶ ｅ前体蛋白体外合成。同时，有２例并未检测到前Ｃ区的变异。结论 部分重型肝炎ＨＢｅＡｇ阴性原因除了Ｔ１８９６变异外，还存在Ｔ１８６２变异。前Ｃ１８６２突变对ＨＢＶｅ抗原前体蛋白合成并无影响，可能ｅ抗原前体在分泌过程中受阻。
Objective To detect mutations in precore region of hepatitis B virus of HBeAg negative - patients with fulminant hepatitis and to determine the effect of T1862 mutants on synthesis of precursor of hepatitis B e antigen. Methods The entire precore and core region were amplified from sera of nine HBeAg negative-patients with fulminant hepatitis B by polymerase chain reaction (PCR). PCR products were cloned into plasmid pUC18, and sequencing for analysis of precore mutations. Precore and core sequence of T1862 variant were also cloned into expression plasmid pGEMT for in vitro transcription and translation study on synthesis and procession of e antigen. Results Three variants, A1896, A1899 and T1862, whose nucleotide mutation led to amino acids substitutions, were detected in patients with fulminant hepatitis. T1862 variant didn't effect the synthesis of precursor of e antigen. Also there was no variant detected in precore region of hepatitis B virus in two patients. Conclusions The causes for negative of e antigen in fulminant hepatitis patients may be partially explained by precore mutation of A1896 and T1862, and the latter variant may effect the process of precursor of e antigen, rather than the synthesis of precursor protein.
出处
《中华肝脏病杂志》
CAS
CSCD
2001年第1期42-44,共3页
Chinese Journal of Hepatology
基金
国家自然科学基金!(39800125)
军队卫生科研课题!(962024)
