DETECTION OF MULTIPLE VIRUSES BY PCR CO-AMPLIFICATION

我们用聚合酶链式反应在一次聚合酶链式反应中同时检测多种病毒。 化验。简单地说，共扩增与来自 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)和HIV-1和乙肝病毒。引子是 在共扩增反应中实现最佳灵敏度的标准化。 用针对一组2个DNA片段的特异性引物对 这些病毒(艾滋病毒-1/艾滋病毒2、艾滋病毒-1/丙型肝炎病毒、艾滋病毒-1/乙肝病毒)及其分析产物 通过与病毒特异性探针杂交。每本都不到10份 用该方法可以检测到病毒DNA。这种检测方法被用来检测 混合感染的临床样本中的各自病毒。我们目前正在 优化共扩增HIV-1/2、乙肝病毒、丙型肝炎病毒和HTLV-I/II基因 用质粒滴定法进行聚合酶链式反应。 描述这项工作的摘要在国际第七届国际会议上提出。电话会议在……上面 艾滋病。佛罗伦萨。已向《华尔街日报》提交了一份手稿 病毒学方法。

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.