Department of Veterinary Public Health and Food Safety, Istituto Superiore di Sanità, Viale Regina Elena 299, 00161 Rome, Italy.
J Virol Methods. 2010 Apr;165(1):57-63. doi: 10.1016/j.jviromet.2010.01.003. Epub 2010 Jan 25.
Viral contamination of drinking water is frequently reported as the primary source of gastroenteritis or hepatitis outbreaks. The presence of viruses at low concentration levels in most environmental water poses major analytical problems when determining their concentration. To evaluate the efficiency of different recovery methods of viral RNA from bottled water, a comparison was made of 2 positively and 2 negatively charged membranes that were used for absorbing and releasing HAV virus particles during the filtration of viral spiked bottled water. All the 4 membranes, regardless of charge and pore size, had low level viral recovery. The results show that a considerable number of the virus particles passed through the pores of the membranes instead of being trapped by the electrostatic charges. Two different procedures were then compared using 1.5L polyethylene bottles spiked with 10-fold serial dilutions of HAV and FCV. The first procedure included an ultrafiltration-based method followed by MiniMag RNA extraction, and the second an ultracentrifugation-based method followed by RNA extraction using QIAamp viral RNA mini kit. The ultracentrifugation-based method resulted in a better recovery of HAV and FCV when compared to the ultrafiltration-based method.
饮用水中的病毒污染经常被报道为肠胃炎或肝炎爆发的主要源头。在大多数环境水中,病毒以低浓度存在,这在确定其浓度时带来了主要的分析问题。为了评估从瓶装水中回收病毒 RNA 的不同方法的效率,对用于在过滤病毒污染的瓶装水时吸附和释放 HAV 病毒颗粒的 2 种正电荷和 2 种负电荷膜进行了比较。所有 4 种膜,无论带电荷和孔径如何,病毒回收率都很低。结果表明,相当数量的病毒颗粒穿过了膜的孔,而不是被静电力捕获。然后使用 1.5L 聚乙烯瓶对 HAV 和 FCV 进行了 10 倍系列稀释,比较了两种不同的程序。第一种程序包括基于超滤的方法,然后是 MiniMag RNA 提取,第二种方法是基于超速离心的方法,然后是使用 QIAamp 病毒 RNA 迷你试剂盒进行 RNA 提取。与基于超滤的方法相比,基于超速离心的方法使 HAV 和 FCV 的回收效果更好。