DETECTION OF HCV SEQUENCES BY USING PCR WITH NESTED PRIMERS

使用带有嵌套引物的 PCR 检测 HCV 序列

• 批准号: 3811116
• 负责人: S NEDJAR
• 金额: --
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/3811116
• 关键词: RNA directed DNA polymerase; blood transfusion; communicable disease control; communicable disease diagnosis; communicable disease transmission; diagnosis design /evaluation; hepatitis C virus; human tissue; nucleic acid sequence; polymerase chain reaction; virus RNA; virus classification; virus protein

Transfusion associated-and so called "community acquired"-hepatitis non-A, non-B is evidently caused by an agent that is, quite distinct hepatitis A and hepatitis B viruses. Recently, a sequence derived from this agent was cloned (Choo, Q.L., et al., Science, 244: 259-262, 1989) . Studies indicate that the agent causing hepatitis C is a positive stranded RNA virus-hepatitis C virus (HCV). At the present time, screening for HCV is based on the presence of antibodies in the sera or plasma of infected individuals to the recombinant HCV peptide, clOO-3, in sera or plasma. This peptide represents a non-structural viral component. Seroconversion (i.e. , the presence of clOO-3 antibody) does not occur for a long period of time (from about six weeks to possibly more than one year) after viral inoculation. Therefore, the feasibility of using PCR to detect HCV-specific sequences during this long window period was investigated. Using PCR in combination with reverse transcription and a pair of nested primers, we have developed a specific and sensitive (to a single copy level) diagnostic method for the detection of HCV sequences in clinical samples.

输血相关的--所谓的“社区获得性”--非甲型肝炎， 非乙型肝炎显然是由一种非常独特的甲型肝炎引起的 和B型肝炎病毒。最近，从这种药剂中获得的序列被 克隆的（Choo，Q.L.，例如，Science，244：259-262，1989）。研究表明 丙型肝炎病毒是一种正链RNA 病毒-丙型肝炎病毒（HCV）。目前，筛查HCV是 基于感染者的血清或血浆中抗体的存在， 个体对血清或血浆中的重组HCV肽c100 -3的敏感性。这 肽代表非结构病毒组分。血清转化（即， cl 00 -3抗体存在）不会长时间发生 (from大约六周到可能超过一年）后，病毒 接种。因此，应用PCR检测HCV特异性抗原的可行性 在这个长的窗口期间的序列进行了研究。PCR在 结合逆转录和一对嵌套引物， 开发了一种特异性和敏感性（到单拷贝水平）的诊断 用于检测临床样品中HCV序列的方法。