Orrù Germano, Masia Giuseppina, Orrù Ginevra, Romanò Luisa, Piras Vincenzo, Coppola Rosa Cristina
Dipartimento di Scienze Odontostomatologiche, Universita' degli Studi di Cagliari, Via Binaghi No. 4, 09121 Cagliari, Italy.
J Virol Methods. 2004 Jun 15;118(2):77-82. doi: 10.1016/j.jviromet.2004.01.025.
Hepatitis E Virus (HEV) is the causative agent of an acute and self-limited form of hepatitis. The virus is transmitted by the faecal-oral route and is a major cause of viral hepatitis in much of the developing world where it causes sporadic infections and large-scale epidemics. A simple and rapid protocol for the measurement of HEV faecal shedding by a real-time polymerase chain reaction (PCR) with the SYBR Green method on a LightCycler instrument, is described. After only 3h the real-time quantitative PCR method detected 10 molecules of HEV cDNA fragment per reaction tube and showed a high linear dynamic range of quantitation (10-10(6) molecules of cDNA/reaction) with a good correlation (r = -1.00). Its specificity was confirmed by assay in human faecal samples.
戊型肝炎病毒（HEV）是一种急性自限性肝炎的病原体。该病毒通过粪-口途径传播，在许多发展中国家是病毒性肝炎的主要病因，可引发散发性感染和大规模流行。本文描述了一种简单快速的方法，用于在LightCycler仪器上采用SYBR Green法通过实时聚合酶链反应（PCR）检测粪便中HEV的排出情况。仅3小时后，实时定量PCR方法就能在每个反应管中检测到10个戊型肝炎病毒cDNA片段分子，并且显示出高线性定量动态范围（10-10⁶个cDNA分子/反应），相关性良好（r = -1.00）。通过在人类粪便样本中的检测证实了其特异性。
