Detection of Noroviruses in Tap Water in Japan by Means of a New Method for Concentrating Enteric Viruses in Large Volumes of Freshwater

Abstract

A virus concentration method using a cation-coated filter was developed for large-volume freshwater applications. Poliovirus type 1 (LSc 2ab Sabin strain) inoculated into 40 ml of MilliQ (ultrapure) water was adsorbed effectively to a negatively charged filter (Millipore HA, 0.45-m pore size) coated with aluminum ions, 99% (range, 81 to 114%) of which were recovered by elution with 1.0 mM NaOH (pH 10.8) following an acid rinse with 0.5 mM H 2 SO 4 (pH 3.0). More than 80% poliovirus recovery yields were obtained from 500-ml, 1,000-ml, and 10-liter MilliQ water samples and from tap water samples. This method, followed by TaqMan PCR detection, was applied to determine the presence of noroviruses in tap water in Tokyo, Japan. In a 14-month survey, 4 (4.1%) and 7 (7.1%) of 98 tap water samples (100 to 532 liters) contained a detectable amount of noroviruses of genotype 1 and genotype 2, respectively. This method was proved to be useful for surveying the occurrence of enteric viruses, including noroviruses, in large volumes of freshwater.

Enteric Viruses of Humans and Animals in Aquatic Environments: Health Risks, Detection, and Potential Water Quality Assessment Tools

“…After the sample is filtered and viruses adsorbed to the membrane, an acid rinse step is used to remove cations (i.e., salt) and other inhibitors while keeping viruses attached to the membrane. In addition, an inorganic eluting medium (NaOH) that has fewer inhibitory effects in PCR assays than the commonly used organic eluting medium (beef extract) is used (69,86); however, a high-pH beef extract solution is the most widely used medium to elute absorbed viruses from cartridge filters and has worked well with cell culture assays (4,136,140). In PCR, the use of NaOH as an eluent provides a good alternative to other methods that attempt to remove PCR inhibitors from beef extract solution, such as resin treatments, polyethylene glycol (PEG) precipitation-resuspension techniques, immunomagnetic capture, and glass purification, which can be expensive and complicated (86).…”

“…For improved recovery of viruses from freshwater, such as groundwater, river water, and tap water, Haramoto et al (69) modified the virus concentration method of Katayama et al (86) by precoating a type HA, negatively charged membrane (Millipore, Billerica, MA) with AlCl 3 prior to filtering samples, yielding a mean poliovirus recovery of 109% from 10 liters of seeded MilliQ water.…”

Detection of Influenza A viruses at migratory bird stopover sites in Michigan, USA

“…Two sample volume ranges were used in this study: 1) small volumes of 300–400 mL and 2) large volumes of 2 L. Negatively-charged 0.45 μm pore size HA filters (Millipore Corporation, Billerica, Massachusetts) of either 47 mm diameter (for 300–400 mL samples) or 90 mm diameter (for 2 L samples) were placed onto a vacuum filter apparatus, and 5 mL of 250 mM AlCl 3 were passed through the membrane to form Al 3 + charged filters [31,32]. Water samples were then vacuum filtered at a maximum filtration rate of 50 mL/min.…”

Adsorption-Elution Techniques and Molecular Detection of Enteric Viruses from Water

“…Electropositive membranes and cartridges are commonly used for the recovery of enteric viruses from water and other materials without prior conditioning of the sample because most enteric viruses are negatively charged at ambient pH [21,22]. The other advantage of the electropositive membranes is the ability to recover rotaviruses (RVs) and coliphages which are sensitive to the low pH needed for electronegative membranes [23].…”