Quantitative assay system established for different hepatitis B virus RNAs in sera sample
- Author:
- No author available
- Journal Title:
- JOURNAL OF THE FOURTH MILITARY MEDICAL UNIVERSITY
- Issue:
- 8
- DOI:
- 10.3321/j.issn:1000-2790.2003.08.001
- Key Word:
- 肝炎病毒,乙型;转录体;癌,肝细胞;血清;聚合酶链反应;拉米夫定;疗效评定
Abstract： 目的: 建立针对血清乙型肝炎病毒(hepatitis B virus, HBV)转录体的定量分析系统.方法: 从血清中提取病毒 DNA 和 RNA,采用竞争性 PCR 技术、Southern-blotting 及分子克隆等技术,建立循环病毒 DNA 和 RNA(转录体)的定量分析系统,并应用其对抗病毒药物 Lamivudine 干预治疗后 HBV 感染患者血清中 HBV DNA 和 RNA 结构和数量的变化进行分析.结果: 建立了两个针对 X 区 RNA 的定量分析系统,可检测带有全长型和顿挫型 3′端结构的转录体.我们还建立了HBV X,Core和X-PreCore区段的 DNA/RNA定量分析系统,三者分别与 HBV 基因复制的早、中、晚期对应.此研究初步阐明了Lamivudine 治疗期间患者血清中 HBV DNA 和转录体结构和数量的变化. 治疗8 wk以后,在Core 和 X-PreCore 区 DNA拷贝数从109*mL-1下降为105*mL-1,下降幅度明显高于 X 区(下降至107*mL-1), 其比率更准确地反映了 Lamivudine 的作用效率,可用于疗效的评估.转录体的拷贝数仅有小幅下降,采用锚定的 Oligo(dT) 引物检测的两种全长和顿挫型Ploy(A) RNA (105*mL-1) 明显低于 X 区 (107*mL-1) RNA 的拷贝数,提示在基因复制过程中前基因组 RNA 的 Ploy(A) 尾被去除. 结论: 这一分析系统可显示血清中 HBV 转录体含量及定量不同3′端结构,为研究循环中 HBV 转录体和 DNA 的动态变化提供了新方法.
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