Biohazard Assessment Research Branch, Microbiological and Chemical Exposure Assessment Research Division, National Exposure Research Laboratory, Office of Research and Development, United States Environmental Protection Agency, 26 West Martin Luther King Drive, Cincinnati, OH 45268, USA.
J Virol Methods. 2013 Oct;193(1):140-6. doi: 10.1016/j.jviromet.2013.05.014. Epub 2013 May 31.
Enteroviruses, noroviruses and adenoviruses are among the most common viruses infecting humans worldwide. These viruses are shed in the feces of infected individuals and can accumulate in wastewater, making wastewater a source of a potentially diverse group of enteric viruses. In this study, two procedures were evaluated to concentrate noroviruses, adenoviruses and enteroviruses from primary effluent of wastewater. In the first procedure, indigenous enteroviruses, noroviruses and adenoviruses were concentrated using celite (diatomaceous earth) followed by centrifugation through a 30K MWCO filter and nucleic acid extraction. The second procedure used celite concentration followed by nucleic acid extraction only. Virus quantities were measured using qPCR. A second set of primary effluent samples were seeded with Coxsackievirus A7, Coxsackievirus B1, poliovirus 1 or enterovirus 70 before concentration and processed through both procedures for recovery evaluation of enterovirus species representatives. The pairing of the single step extraction procedure with the celite concentration process resulted in 47-98% recovery of examined viruses, while the celite concentration process plus additional centrifugal concentration before nucleic acid extraction showed reduced recovery (14-47%). The celite concentration process followed by a large volume nucleic acid extraction technique proved to be an effective procedure for recovering these important human pathogens from wastewater.
肠道病毒、诺如病毒和腺病毒是全球范围内感染人类最常见的病毒之一。这些病毒存在于受感染个体的粪便中,并可在废水中积聚,使废水成为潜在多种肠道病毒的来源。在这项研究中,评估了两种程序来浓缩废水中的原生肠道病毒、诺如病毒和腺病毒。在第一个程序中,使用硅藻土(硅藻土)浓缩土着肠道病毒、诺如病毒和腺病毒,然后通过 30K MWCO 过滤器离心和核酸提取。第二个程序仅使用硅藻土浓缩,然后进行核酸提取。使用 qPCR 测量病毒数量。在浓缩前,将第二批原初废水样本接种柯萨奇病毒 A7、柯萨奇病毒 B1、脊髓灰质炎病毒 1 或肠道病毒 70,然后通过两种程序进行处理,以评估肠道病毒种代表的回收情况。将单步提取程序与硅藻土浓缩过程结合使用,可使检测到的病毒回收率达到 47-98%,而在核酸提取前增加离心浓缩的硅藻土浓缩过程显示回收率降低(14-47%)。事实证明,硅藻土浓缩过程后采用大体积核酸提取技术是从废水中回收这些重要人类病原体的有效程序。