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Applied and Environmental Microbiology, March 2002, p. 1033-1039, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1033-1039.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.

Development of a Virus Concentration Method and Its Application to Detection of Enterovirus and Norwalk Virus from Coastal Seawater

Hiroyuki Katayama,* Akihiro Shimasaki, and Shinichiro Ohgaki

Department of Urban Engineering, School of Engineering, University of Tokyo, Bunkyo-ku, Tokyo 113-8656, Japan

Received 5 July 2001/ Accepted 11 December 2001

We developed a new procedure for concentration of enteric viruses from water using a negatively charged membrane. Rinsing the membrane with 0.5 mM H2SO4 (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.

* Corresponding author. Mailing address: Department of Urban Engineering, School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. Phone: 81-3-5841-6245. Fax: 81-3-5841-8533. E-mail: katayama{at}env.t.u-tokyo.ac.jp.

Applied and Environmental Microbiology, March 2002, p. 1033-1039, Vol. 68, No. 3
0099-2240/02/$04.00+0     DOI: 10.1128/AEM.68.3.1033-1039.2002
Copyright © 2002, American Society for Microbiology. All Rights Reserved.



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