MOLECULAR BIOLOGY OF HEPATITIS C VIRUS

丙型肝炎病毒(丙型肝炎病毒)是一种重要的人类病原体 与输血相关的非甲非乙型(NANB)肝炎有关。 逆转录聚合酶扩增丙型肝炎病毒RNA序列 链式反应(Cdna)是目前唯一实用的方法。 目的：证实丙型肝炎病毒感染者的病毒血症。然而，遗传的 不同丙型肝炎病毒株间的异质性导致假阴性结果 在cDNAPCR分析中，由于引物和模板的不匹配。丙型肝炎病毒账户 对于高达25%的社区获得性肝炎和90%以上的输血- 美国的关联性肝炎。最近的克隆和 丙型肝炎病毒基因组测序导致特异性 丙型肝炎病毒抗体(抗-丙型肝炎病毒)的血清学检测 衍生的病毒抗原。这些测试对诊断和治疗非常有用 筛查的目的，但它们在一定程度上没有揭示丙型肝炎病毒的免疫力 因为目前可用的抗原是从部分丙型肝炎病毒表达出来的 编码非结构或内部结构蛋白的基因组。 此外，发现丙型肝炎病毒基因组的遗传异质性， 特别是在编码包膜蛋白的基因中，表明 可能是病毒包膜蛋白的异质性，类似于 人类免疫缺陷病毒。这样的发现对尝试来说不是好兆头 在疫苗研发方面。本项目的目标是：(1) 鉴定c DNA聚合酶链式反应中假阴性率低的引物 检测；(2)确定不同丙型肝炎病毒基因组的核苷酸序列；以及 (3)分析灵长类动物感染丙型肝炎病毒是否诱导保护性 豁免权。

Hepatitis C virus (HCV) is an important human pathogen that is strongly associated with transfusion related non-A, non-B (NANB) hepatitis. Amplification of HCV RNA sequences by reverse transcription and polymerase chain reaction (cDNA PCR) is the only practical method currently available to demonstrate viremia in patients with HCV infection. However, genetic heterogeneity among different HCV strains results in false negative results in cDNA PCR assays because of primer and template mismatch. HCV accounts for up to 25% of community-acquired hepatitis and over 90% of transfusion- associated hepatitis in the United States. The recent cloning and sequencing of the genome of HCV led to the development of specific serologic tests for antibody to HCV (anti-HCV) that utilized recombinant- derived viral antigens. These tests have been useful for diagnostic and screening purposes but they shed little light on immunity to HCV, in part because currently available antigens are expressed from parts of the HCV genome that encode nonstructural or internal structural proteins. Furthermore, the finding of genetic heterogeneity of the HCV genome, especially in the gene encoding the envelope proteins, suggests that there may be heterogeneity of viral envelope proteins similar to that seen in human immunodeficiency viruses. Such a finding would bode ill for attempts at vaccine development. The objectives of this project are to: (1) identify primer sets that have a low false negative rate in a cDNA PCR assay; (2) determine the nucleotide sequence of diverse HCV genomes; and (3) analyze whether infection of primates with HCV induces protective immunity.