Short communication
A novel method for concentrating hepatitis A virus and caliciviruses from bottled water

https://doi.org/10.1016/j.jviromet.2009.07.013Get rights and content

Abstract

Human enteric viruses are detected frequently in various types of environmental water samples, such as irrigation water, wastewater, recreational water, ground or subsurface water and even drinking water, constituting a primary source of gastroenteritis or hepatitis outbreaks. Only a few, but still infective number of viral particles are normally present in water samples, therefore an efficient virus concentration procedure is essential prior to molecular detection of the viral nucleic acid. In this study, a novel chromatographic technology, Convective Interaction Media® (CIM) monolithic supports, were optimized and applied to the concentration of hepatitis A virus (HAV) and feline calicivirus (FCV), a surrogate of norovirus (NoV), from water samples. Two-step real-time RT-qPCR was used for quantitation of the virus concentration in the chromatographic fractions. Positively charged CIM QA (quaternary amine) monolithic columns were used for binding of HAV and FCV present in previously inoculated 1.5 l bottled water samples. Column bound viruses were eluted from the monolith using 1 M NaCl to a final volume of 15 ml. Elution volume was concentrated further by ultracentrifugation. When the CIM/ultracentrifugation method was compared with another concentration method employing positively charged membranes and ultrafiltration, the recovery of HAV was improved by approximately 20%.

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Acknowledgements

We thank Dr. Dario De Medici and Dr. Simona Di Pasquale (Istituto Superiore Di Sanità, Rome, Italy) who kindly provided stocks of hepatitis A virus and feline calicivirus and also primers, probes and real-time RT-PCR protocols for both types of viruses.

This work was financed by BIOTRACER (Project No. 036272) which is an integrated project within the European Union 6th framework programme under the European research area of food quality and safety with full name Improved bio-traceability of

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