Key result
A new virus concentration method using a negatively charged membrane with an acid rinse and alkaline elution achieved 89% recovery of poliovirus from natural seawater, compared to 38% without the acid rinse.
A novel virus concentration method using a negatively charged membrane with an acid rinse significantly improves the recovery and detection of enteric viruses from seawater compared to conventional methods.
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May enhance enteric virus surveillance in seawater; leaves open validation for routine environmental monitoring.
We developed a new procedure for concentration of enteric viruses from water using a negatively charged membrane. Rinsing the membrane with 0.5 mM H(2)SO(4) (pH 3.0) in order to elute cations prior to viral elution with 1 mM NaOH (pH 10.5) promoted poliovirus recovery yields from 33 to 95% when applied to pure water and 38 to 89% when applied to natural seawater from Tokyo Bay, Japan, respectively. This method showed average recovery yields of spiked poliovirus of 62% (n = 8) from 1 liter of artificial seawater. This method showed higher recovery yields (>61%) than that of the conventional method using positively charged membrane (6%) when applied to seawater. This method is also free from beef extract elution, which has an inhibitory effect in the subsequent viral genome detection by reverse transcription-PCR. Naturally occurring Norwalk viruses from 2 liters of Tokyo Bay water in winter and infectious enteroviruses from 2 liters of recreational coastal seawater in summer were detected by using this viral concentration method.
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Katayama et al. (2002) studied Viral contamination of seawater. Negatively charged membrane with acid rinse and alkaline elution vs. Conventional method without acid rinse was evaluated on Poliovirus recovery yield from natural seawater. A new virus concentration method using a negatively charged membrane with an acid rinse and alkaline elution achieved 89% recovery of poliovirus from natural seawater, compared to 38% without the acid rinse.
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