Gibellini Davide, Gardini Federica, Vitone Francesca, Schiavone Pasqua, Furlini Giuliano, Re Maria Carla
Department of Clinical and Experimental Medicine, Microbiology Section, University of Bologna, St. Orsola Hospital, Via Massarenti 9, 40138 Bologna, Italy.
Mol Cell Probes. 2006 Jun-Aug;20(3-4):223-9. doi: 10.1016/j.mcp.2005.12.005. Epub 2006 Mar 14.
This paper describes the development of a SYBR Green-based multiplex real time RT-PCR for the simultaneous detection of HCV and HIV-1 genomes in plasma samples. Viral genomes were identified in the same sample by their distinctive melting temperature (Tm) which are 81.6 and 86.5 degrees C for HIV-1 gag 142 bp amplicon and HCV 5'-NCR region 226 bp amplicon, respectively. Analysis of known scalar concentrations of reference plasma indicated that the multiplex procedure detects at least 500 copies/ml of both HIV-1 and HCV. In addition, we also assayed HIV-1 and HCV viral load in 30 co-infected patients and in 15 blood donors, confirming the sensitivity and specificity of the assay. This method may represent a useful alternative method for the detection of HIV-1/HCV co-infection, reliable for a rapid and relatively inexpensive screening of blood donors.
本文描述了一种基于SYBR Green的多重实时逆转录聚合酶链反应（RT-PCR）的开发，用于同时检测血浆样本中的丙型肝炎病毒（HCV）和人类免疫缺陷病毒1型（HIV-1）基因组。通过其独特的解链温度（Tm）在同一样本中鉴定病毒基因组，HIV-1 gag 142 bp扩增子和HCV 5'-非编码区（NCR）226 bp扩增子的解链温度分别为81.6和86.5摄氏度。对已知标量浓度的参考血浆进行分析表明，多重检测程序可检测到至少500拷贝/毫升的HIV-1和HCV。此外，我们还检测了30例合并感染患者和15例献血者的HIV-1和HCV病毒载量，证实了该检测方法的敏感性和特异性。该方法可能是检测HIV-1/HCV合并感染的一种有用的替代方法，对于快速且相对廉价地筛查献血者是可靠的。
