Park Geun Woo, Chhabra Preeti, Vinjé Jan
Division of Viral Diseases, Centers for Disease Control and Prevention;
Division of Viral Diseases, Centers for Disease Control and Prevention.
J Vis Exp. 2017 Feb 6(120):55205. doi: 10.3791/55205.
Human noroviruses are a leading cause of epidemic and sporadic gastroenteritis worldwide. Because most infections are either spread directly via the person-to-person route or indirectly through environmental surfaces or food, contaminated fomites and inanimate surfaces are important vehicles for the spread of the virus during norovirus outbreaks. We developed and evaluated a protocol using macrofoam swabs for the detection and typing of human noroviruses from hard surfaces. Compared with fiber-tipped swabs or antistatic wipes, macrofoam swabs allow virus recovery (range 1.2-33.6%) from toilet seat surfaces of up to 700 cm. The protocol includes steps for the extraction of the virus from the swabs and further concentration of the viral RNA using spin columns. In total, 127 (58.5%) of 217 swab samples that had been collected from surfaces in cruise ships and long-term care facilities where norovirus gastroenteritis had been reported tested positive for GII norovirus by RT-qPCR. Of these 29 (22.8%) could be successfully genotyped. In conclusion, detection of norovirus on environmental surfaces using the protocol we developed may assist in determining the level of environmental contamination during outbreaks as well as detection of virus when clinical samples are not available; it may also facilitate monitoring of effectiveness of remediation strategies.
人诺如病毒是全球流行性和散发性肠胃炎的主要病因。由于大多数感染要么通过人际传播途径直接传播，要么通过环境表面或食物间接传播，受污染的污染物和无生命表面是诺如病毒爆发期间病毒传播的重要载体。我们开发并评估了一种使用大泡沫拭子从硬表面检测和分型人诺如病毒的方案。与纤维头拭子或防静电擦拭布相比，大泡沫拭子可从长达700厘米的马桶座圈表面回收病毒（回收率为1.2%-33.6%）。该方案包括从拭子中提取病毒以及使用旋转柱进一步浓缩病毒RNA的步骤。在从报告了诺如病毒肠胃炎的游轮和长期护理机构的表面收集的217个拭子样本中，共有127个（58.5%）通过RT-qPCR检测出GII诺如病毒呈阳性。其中29个（22.8%）能够成功进行基因分型。总之，使用我们开发的方案检测环境表面的诺如病毒可能有助于确定疫情期间的环境污染水平以及在没有临床样本时检测病毒；它还可能有助于监测补救策略的有效性。
