Two-Step Concentration of Complex Water Samples for the Detection of Viruses

Methods and Protocols

Jones, Davey L.¹  Farkas, Kata²  McDonald, James E.³  Malham, Shelagh K.⁴ 

[1] Author to whom correspondence should be addressed.;School of Natural Sciences, Bangor University, Bangor LL57 2UW, UK;School of Ocean Sciences, Bangor University, Menai Bridge LL59 5AB, UK;UWA School of Agriculture and Environment, University of Western Australia, Crawley 6009, Australia

关键词: tangential flow ultrafiltration;    virus precipitation;    qPCR;    enteric viruses;    norovirus;    mengovirus;   

DOI  :  10.3390/mps1030035

学科分类：生物技术

来源: mdpi

【 摘 要 】

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.

【 授权许可】

CC BY