Farkas Kata, McDonald James E, Malham Shelagh K, Jones Davey L
School of Natural Sciences, Bangor University, Bangor LL57 2UW, UK.
School of Ocean Sciences, Bangor University, Menai Bridge LL59 5AB, UK.
Methods Protoc. 2018 Sep 10;1(3):35. doi: 10.3390/mps1030035.
The accurate detection and quantification of pathogenic viruses in water is essential to understand and reduce the risk of human infection. This paper describes a two-step method suitable for concentrating viruses in water and wastewater samples. The method involves a tangential flow ultrafiltration step that reduces the sample volume of 1-10 L to approximately 50 mL, followed by secondary precipitation using polyethylene glycol 6000, which reduces the volume to 1-4 mL. For method validation, water samples were spiked with different concentrations of enteric viruses, and viral recovery in the concentrates exceeded 10% in all experiments. The method is suitable for water samples with high and low salinity and turbidity, allowing an accurate comparison of viral titers in a diverse range of water types. Furthermore, the method has the potential to concentrate other pathogens, e.g., bacteria or protozoa. Hence, the use of this method can improve the holistic assessment of risks associated with wastewater-contaminated environments.
准确检测和定量水中的致病病毒对于了解和降低人类感染风险至关重要。本文描述了一种适用于浓缩水和废水样本中病毒的两步法。该方法包括一个切向流超滤步骤，可将1-10 L的样本体积减少至约50 mL，随后使用聚乙二醇6000进行二次沉淀，将体积减少至1-4 mL。为验证该方法，向水样中加入不同浓度的肠道病毒，在所有实验中浓缩物中的病毒回收率均超过10%。该方法适用于高盐度、低盐度和不同浊度的水样，能够准确比较各种不同类型水中的病毒滴度。此外，该方法有潜力浓缩其他病原体，如细菌或原生动物。因此，使用该方法可以改善对受废水污染环境相关风险的整体评估。
