Sherker A H, Twu J S, Reyes G R, Robinson W S
Division of Infectious Diseases, Stanford University School of Medicine, California 94305.
J Med Virol. 1993 Feb;39(2):91-6. doi: 10.1002/jmv.1890390203.
Hepatitis C virus (HCV) is a positive-polarity, single-stranded RNA virus, distantly related to the pestivirus and flavivirus genera. These viruses replicate through the formation of a minus-strand RNA intermediate, which encodes the positive-strand genome, which is subsequently encapsidated, enveloped, and released from infected cells. Minus-strand RNA is not found in the mature, circulating virions of flaviviruses. In an attempt to study the relative amounts of viral plus and minus strand in the liver and serum of HCV-infected individuals, we have developed a technique to amplify specifically each of the viral strands using a modified reverse transcriptase/polymerase chain reaction protocol on extracted RNA. Liver tumor and nontumor tissue from a patient with C-100-3 antibody was analyzed using this technique. In both cases, viral plus and minus strands were detected, although the plus-strand signal was several fold stronger than minus-strand signal by Southern hybridization. Sera from 11 C-100-3 antibody-positive patients with abnormal serum AIT levels were similarly analyzed. In all cases viral plus strand was detected, and in 10 of 11 cases viral minus strand was detected. The minus-strand signal was always much weaker than the plus-strand signal and the ratio of plus strand to minus strand varied among patients. No correlation was found between the level of minus strand detected or its ratio to plus strand with the level of serum transaminases or any other clinical parameter.
丙型肝炎病毒（HCV）是一种正链单链RNA病毒，与瘟病毒属和黄病毒属有较远的亲缘关系。这些病毒通过形成负链RNA中间体进行复制，该中间体编码正链基因组，随后该基因组被衣壳化、包膜化并从受感染细胞中释放出来。在黄病毒成熟的循环病毒粒子中未发现负链RNA。为了研究丙型肝炎病毒感染个体肝脏和血清中病毒正链和负链的相对含量，我们开发了一种技术，使用改良的逆转录酶/聚合酶链反应方案对提取的RNA进行特异性扩增，以扩增病毒的每条链。使用该技术分析了一名C-100-3抗体阳性患者的肝肿瘤和非肿瘤组织。在这两种情况下，均检测到病毒正链和负链，尽管通过Southern杂交检测到的正链信号比负链信号强几倍。对11名血清谷丙转氨酶（AIT）水平异常的C-100-3抗体阳性患者的血清进行了类似分析。在所有病例中均检测到病毒正链，11例中有10例检测到病毒负链。负链信号总是比正链信号弱得多，且正链与负链的比例在患者之间有所不同。未发现检测到的负链水平或其与正链的比例与血清转氨酶水平或任何其他临床参数之间存在相关性。
