Komurian-Pradel Florence, Perret Magali, Deiman Birgit, Sodoyer Mireille, Lotteau Vincent, Paranhos-Baccalà Glaucia, André Patrice
UMR2142 CNRS-bioMérieux, IFR128 Biosciences Lyon-Gerland, 21 Avenue, Tony Garnier, 69365, Lyon Cedex 07, France.
J Virol Methods. 2004 Mar 1;116(1):103-6. doi: 10.1016/j.jviromet.2003.10.004.
Qualitative detection of negative hepatitis C virus (HCV) RNA has been used widely to demonstrate HCV replication. However, relative quantitation of both positive and negative HCV RNA strands has never been reported for studying viral genome replication. A strand specific real-time PCR carried out in the highly conserved 5'-non-coding region of HCV genome and monitored either by the DNA binding dye SYBR Green I or by molecular beacons is described. Using these techniques, it was found that negative HCV RNA strand was a 100-1000 times less abundant than the positive strand in the liver of HCV infected patients.
丙型肝炎病毒（HCV）RNA阴性的定性检测已被广泛用于证明HCV复制。然而，尚未有关于研究病毒基因组复制的HCV正负链RNA相对定量的报道。本文描述了一种在HCV基因组高度保守的5'-非编码区进行的链特异性实时PCR，该PCR通过DNA结合染料SYBR Green I或分子信标进行监测。使用这些技术发现，在HCV感染患者的肝脏中，HCV RNA负链的丰度比正链低100-1000倍。
