NON-A NON-B HEPATITIS

非甲非乙型肝炎(NANBH)自 霍顿成功克隆丙型肝炎病毒基因组 等人的研究。在奇龙。我们开发了一种敏感的基于聚合酶链式反应的方法来检测 临床样本中的病毒基因组。这个测试似乎是原来的10倍。 比黑猩猩更敏感的传染性。我们已经展示了它的 检测40例慢性NANBH患者血清的有效性。 有趣的是，在36名患者中，只有16人的丙型肝炎病毒抗体呈阳性。 用聚合酶链式反应检测血清中的丙型肝炎病毒核糖核酸。作为 检测的灵敏度很高，这一点尚不清楚 为什么阳性率这么低。克隆和测序揭示了 丙型肝炎病毒毒株之间高度的序列多样性。vbl.使用 我们现在已经在这40个人中的35个人中检测到了丙型肝炎病毒RNA 患者，包括所有4名在商业广告中未检测到抗体的患者 测试。最近我们发现丙型肝炎病毒RNA的5‘非编码区 菌株间基因组高度保守。我们现在准备好了聚合酶链式反应 这一地区的引物似乎是通用的。 我们已经克隆和测序了丙型肝炎病毒基因组的主要部分，包括 大约2000个碱基，代表基因组的5‘端， 以前没有出版过(现在已经在日本出版了)。 这是丙型肝炎病毒结构基因的编码区。这一地区 看起来很独特，比同一地区短得多 典型的黄病毒。我们已经从这个开始进行了表达实验 希望能生产出有用的抗原。 我们正在用Charles表达基因组中可能的蛋白水解酶区域 以大米为研究对象，研究病毒粒子多蛋白的加工工艺及大米中的 酶本身的结构和功能。 我们正在开发可能的组织培养系统，与 斯坦福大学的哈里·格林伯格和LCI的柯蒂斯·哈里斯。这些实验是 以人胎肝和连续的成年人和黑猩猩为基础 细胞系。

Progress in non-A,non-B hepatitis (NANBH) has been rapid since the successful cDNA cloning of the hepatitis C virus (HCV) genome by Houghton et al. at Chiron. We have developed a sensitive PCR based method to detect the viral genome in clinical samples. This assay seems to be about 10 fold more sensitive that chimpanzee infectivity. We have demonstrated its usefulness by testing sera from 40 patients with chronic NANBH. Interestingly, only 16 of patients 36 of whom were positive for anti-HCV by the Chiron assay had HCV RNA in their serum as measured by PCR. As the sensitivity of the assay was shown to be very high, it was not understood why the positivity rate was so low. Cloning and sequencing has revealed a very high degree of sequence diversity between strains of HCV. Using multiple primer sets we have now detected HCV RNA in 35 of these 40 patients including all 4 who had no detectible antibody with the commercial test. Recently we have shown that the 5' non-coding region of the HCV RNA genome is highly conserved between strains. we have now prepared PCR primers from this region that appear to be universal. We have cloned and sequenced major portions of the genome of HCV including approximately 2000 bases representing the 5' terminus of the genome that had not previously been published (it has now been published in Japan). This is the coding region for the structural genes of HCV. This region seems to be quite unique and is much shorter than the same region in typical flaviviruses. We have initiated expression experiments from this region with the hope of producing useful antigens. We are expressing the putative protease region of the genome with Charles Rice in order to study the processing of the virion polyprotein and the structure and function of the enzyme itself. We are developing possible tissue culture systems in collaboration with Harry Greenberg at Stanford and Curtis Harris at LCI. These experiments are based on human fetal liver and on continuous adult human and chimpanzee cell lines.