Analysis of Virus Genome in Sera from Patients with Silent Hepatitis B and Establishment of Detection Method for Antibodies against Virus

我们研究了10例急性肝炎，其中包括2例非药物、酒精或自身免疫性的重型肝炎，其中甲-戊型肝炎病毒标志物为阴性。10例患者血清中5‘端非翻译区和解旋酶区域均未扩增出庚型肝炎病毒基因组。10例患者中有8例(包括2例重型肝炎)前C区套式聚合酶链式反应扩增出乙肝病毒(HBVDNA)。此外，我们还研究了另外5例甲型-戊型肝炎病毒标志物阴性的重型肝炎患者。套式聚合酶链式反应的5例患者中有4例在同一区域扩增出HBVDNA。用直接测序法对扩增产物进行分析。在6例重型肝炎患者中，有4例发现前C区28位密码子(TGG)突变为终止密码子(TAG)。6例重型肝炎患者中3例前S2区扩增，X区未扩增出HBVDNA。我们以前C区和前S2区为立足点，尝试用引物走步的方法对未知残留区进行序列分析。然而，到目前为止，我们还没有得到有用的结果来建立针对沉默的乙肝病毒抗体的检测方法。

We studied 10 patients with acute hepatitis, including 2 cases of fulminant hepatitis, that was not drug-induced, alcoholic, or autoimmune and in which viral markers for hepatitis A-E were negative. Hepatitis G virus genome was not amplified in sera obtained from the 10 patients with RT-PCR at 5'-untranslted and helicase regions. Hepatits B virus (HBV) genome was amplified in 8 of the 10 patients, including 2 cases of fulminant hepatitis, with nested PCR at the precore region. Futhermore, we studied other 5 patients with fulminant hepatitis, in which viral markers for hepatitis A-E were negative. HBV genome was also amplified in 4 of the 5 patients with nested PCR at the same region. Amplified PCR products were analysed with the direct sequencing method. The point mutation that changes codon 28 (TGG) of the precore region to stop codon (TAG) was recognized in 4 of the 6 patiedts with fulminant hepatitis. Although HBV genome was not amplified with nested PCR at the X region in all patients,but amplified at the preS2 region in 3 of 6 patients with fulminant hepatitis. By using the precore and preS2 regions as a foothold, we tried to analyse the sequence of residual unknown region with the primer walking method. However, we cannot get the useful results so far to establish the detecction method for antibodies against the silent HBV.