Presentations

Document Type

Poster

Publication Date

2008

Department

Biological Sciences

Abstract

Viruses such as Rotavirus, Adenovirus and Norovirus are important etiological agents of gastroenteritis worldwide. With the high sensitivity and specificity of PCR, it is now possible to develop PCR-based methods to detect and quantify pathogenic viruses in environmental water samples. To develop reliable methods however, an effective procedure to concentrate viruses from large volumes of water is required. Because of the scale of concentration required, the procedure often requires two steps. The first to reduce tens of liters of water to less than half a liter and then a second to concentrate the sample to a final volume of less than 10 mL for RNA/DNA extraction. The objectives of the study were to compare the efficacy of hollow fiber ultrafiltration (HFUF) using F200B to that of an adsorption/elution method (AEM) using positively charged filters for concentrating viruses for the first step and to compare polyethylene glycol (PEG) precipitation to centrifugal ultrafiltration for the second step. A third objective was to determine the viral detection limit using real-time RT-PCR. Using beach water spiked with a singlestranded RNA bacteriophage (MS2) as a model, our results show a virus recovery rate of 84±6% and 18±8% for the HFUF method and AEM, respectively. For the second concentration step, we obtained a recovery rate of 49±5 % and 87±7% using PEG precipitation and centrifugal ultrafiltration, respectively. A potential limiting factor to more widespread using of HFUF is the higher cost and we found that cost can be reduced by using reusable filters. We were able to sanitize and reuse the same filter at least six times without affecting the virus recovery rate or the processing time.

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