DETECTION OF MULTIPLE VIRUSES BY PCR CO-AMPLIFICATION

PCR 联合扩增检测多种病毒

• 批准号: 3804838
• 负责人: I K HEWLETT
• 金额: --
• 依托单位: CENTER BIOLOGICS EVALUATION RESEARCH TRANSFUSION
• 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/3804838
• 关键词: communicable disease diagnosis; diagnosis design /evaluation; diagnostic tests; hepatitis B virus group; human T cell lymphotropic virus type 1; human T cell lymphotropic virus type 2; human immunodeficiency virus 1; human immunodeficiency virus 2; immunopathology diagnosis; nucleic acid hybridization; nucleic acid probes; nucleic acid sequence; plasmids; polymerase chain reaction; virus DNA

We have used PCR to simultaneously detect multiple viruses in a single PCR assay. Briefly, co-amplification was performed with primers derived from the gag region of HIV-1 (SK 38/39) and the LTR region of HIV-2 (SK 89/90), HIV-1 and HCV (hepatitis C virus) and HIV-1 and HBV. Primers were standardized for optimal sensitivity in the co-amplification reaction. Amplification was performed with primer pairs specific for a set of 2 of these viruses (HIV-1/HIV 2, HIV-1/HCV, HIV-1/HBV) and the products analyzed by hybridization with virus specific probes. Fewer than 10 copies of each viral DNA could be detected by this method. This assay was used to detect the respective viruses in co-infected clinical samples. We are currently optimizing co-amplify HIV-1/2, HBV, HCV and HTLV-I/II sequences in a single PCR assay using plasmid titration. Abstracts describing this work were presented at the VII Int. Conf. on AIDS. Florence. A manuscript has been submitted to the Journal of Virological Methods.

我们已经使用PCR在一个PCR中同时检测多种病毒 比色法 简而言之，共扩增用来源于以下的引物进行： HIV-1的gag区（SK 38/39）和HIV-2的LTR区（SK 89/90）， HIV-1和HCV（丙型肝炎病毒）以及HIV-1和HBV。 引物 标准化以用于共扩增反应中的最佳灵敏度。 用特异于一组2个引物的引物对进行扩增。 这些病毒（HIV-1/HIV 2、HIV-1/HCV、HIV-1/HBV）和分析的产物 通过与病毒特异性探针杂交。 每种都少于10份 该方法可检测到病毒DNA。 该试验用于检测 共感染临床样本中的相应病毒。 我们目前正在 优化在单一扩增中共扩增HIV-1/2、HBV、HCV和HTLV-I/II序列， 使用质粒滴定的PCR测定。 描述这项工作的摘要已在第七届国际会议上提交。 艾滋病 佛罗伦萨。 一份手稿已提交给《华尔街日报》， 病毒学方法。