Manzin A, Solforosi L, Bianchi D, Gabrielli A, Giostra F, Bruno S, Clementi M
Istituto di Microbiologia, Università di Ancona, Italy.
Res Virol. 1995 Jul-Aug;146(4):279-84. doi: 10.1016/0923-2516(96)80572-3.
Molecular methods for the absolute quantitation of nucleic acids present in biological samples have recently been developed and applied in basic and in medical virology; these studies indicated that competitive polymerase chain reaction (PCR) and competitive reverse transcription PCR (cRT-PCR)-based methodologies are currently the methods of choice for quantifying DNA and RNA species present in clinical samples at low concentration. Recently, quantitative molecular techniques were developed to study the hepatitis C virus (HCV) pathogenic potential, the natural history of HCV-infected patients and the efficiency of antiviral therapies in real time. The pilot study reported here was carried out using a cRT-PCR application for the direct quantitation of HCV RNA molecules in plasma samples of infected individuals which was recently developed in our laboratory. Although sharp individual variability of viral load was documented in this study, the mean HCV RNA copy number detected in samples from untreated HCV-infected patients with various clinical conditions (chronic active hepatitis, cirrhosis, cryoglobulinaemia and chronic hepatitis) was substantially similar, with only one exception: in samples from patients tested positive for anti-liver-kidney microsomal (anti LKM1) auto-antibodies, a significantly lower HCV viraemia level was revealed. Additionally, HCV viraemia was monitored in four patients with sustained biochemical and histological response (at least 12 months) following interferon-alpha discontinuation.
用于绝对定量生物样品中核酸的分子方法最近已被开发并应用于基础和医学病毒学领域;这些研究表明,基于竞争性聚合酶链反应(PCR)和竞争性逆转录PCR(cRT-PCR)的方法目前是定量临床样品中低浓度DNA和RNA种类的首选方法。最近,开发了定量分子技术来实时研究丙型肝炎病毒(HCV)的致病潜力、HCV感染患者的自然病史以及抗病毒治疗的效果。本文报道的初步研究是使用cRT-PCR应用进行的,该应用用于直接定量感染个体血浆样品中的HCV RNA分子,这是我们实验室最近开发的。尽管本研究记录了病毒载量的明显个体差异,但在患有各种临床病症(慢性活动性肝炎、肝硬化、冷球蛋白血症和慢性肝炎)的未经治疗的HCV感染患者的样品中检测到的平均HCV RNA拷贝数基本相似,只有一个例外:在抗肝肾微粒体(抗LKM1)自身抗体检测呈阳性的患者样品中,HCV病毒血症水平明显较低。此外,对4例在停用α干扰素后具有持续生化和组织学反应(至少12个月)的患者进行了HCV病毒血症监测。