Prevention of waterborne disease outbreaks relies on the efficient detection of pathogens in drinking and recreational water. Development of sample concentration technology that ensures fast and high recovery of pathogens from aquatic samples is crucial for reliable detection. EPA-approved method VIRADEL (VIRus ADsorption and Elution) is expensive, exhibits low and poorly reproducible recoveries for certain viruses such as adenoviruses. Crossflow ultrafiltration (UF) is an emerging alternative method of waterborne pathogen concentration. The membrane with an appropriate pore size can simultaneously concentrate a wide range of microbial pathogens without concentrating molecular toxins and low molecular weight qPCR inhibitors. To decrease virus adhesion to the membrane, UF membranes are coated with calf serum. In contrast to VIRADEL, UF technique exhibits significantly higher recovery for some viruses. However, overnight deposition of calf serum on UF membranes is a poorly controlled and time consuming process. Such protein coating is also prone to contamination during transport and under field sampling conditions.To overcome this limitation, we propose a conceptually novel approach to virus concentration based on sacrificial membrane films. Such films can be prepared by coating an ultrafiltration membrane with a polyelectrolyte multilayer (PEM) using layer-by-layer adsorption of polycations and polyanions. Main advantages of the proposed approach are (1) fast and reproducible preparation of membranes (less than 1 hour), (2) highly controllable properties (charge, hydrophilicity, swellability, etc) resulting in a highly anti-adhesive membrane surface, (3) dissolution of coatings to recover any virus that is adsorbed on the film.We designed anti-adhesive sacrificial membrane films by coating polyethersulfone membrane with a PEM consisting of alternating layers of chitosan and heparin and assessed the effectiveness of the coated membrane in recovering human adenovirus 40 (HAdV 40), a non-enveloped, double stranded DNA virus that is responsible for most cases of adenovirus-associated gastroenteritis. We also comparatively evaluated the recovery of such virus by traditional calf serum blocked membrane. To our knowledge, this is the first study that evaluates both of these approaches for HAdV recovery from water. Our results demonstrate that coating a membrane with a PEM film results in very high pre-elution recovery from DI water (~90%). HAdV 40 recovery from surface water is ~60 % for both PEM-coated and calf serum-blocked membranes.

Novel membranes for adenovirus recovery from water

248th National Meeting (2014)

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