Development of a virus concentration method and its application to detection of enterovirus and norwalk virus from coastal seawater.
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.
Katayama
Hiroyuki
H
Department of Urban Engineering, School of Engineering, University of Tokyo, 7-3-1 Hongo, Bunkyo-ku, Tokyo 113-8656, Japan. katayama@env.t.u-tokyo.ac.jp
Shimasaki
Akihiro
A
Ohgaki
Shinichiro
S
D004770:Enterovirus / Q000302:isolation & purification*; D009663:Norwalk virus / Q000302:isolation & purification*; D012623:Seawater; D014773:Virology / Q000379:methods; D014871:Water Microbiology