title: Microfluidic quantitative PCR for simultaneous quantification of multiple viruses in environmental water samples.
abstract: To secure food and water safety, quantitative information on multiple pathogens is important. In this study, we developed a microfluidic quantitative PCR (MFQPCR) system to simultaneously quantify 11 major human viral pathogens, including adenovirus, Aichi virus, astrovirus, enterovirus, human norovirus, rotavirus, sapovirus, and hepatitis A and E viruses. Murine norovirus and mengovirus were also quantified in our MFQPCR system as a sample processing control and an internal amplification control, respectively. River water contaminated with effluents from a wastewater treatment plant in Sapporo, Japan, was collected and used to validate our MFQPCR system for multiple viruses. High-throughput quantitative information was obtained with a quantification limit of 2 copies/μl of cDNA/DNA. Using this MFQPCR system, we could simultaneously quantify multiple viral pathogens in environmental water samples. The viral quantities obtained using MFQPCR were similar to those determined by conventional quantitative PCR. Thus, the MFQPCR system developed in this study can provide direct and quantitative information for viral pathogens, which is essential for risk assessments.
authors.affiliation: Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan s.ishii@eng.hokudai.ac.jp.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Ishii
authors.affiliation: Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan.
authors.forename: Gaku
authors.initials: G
authors.lastname: Kitamura
authors.affiliation: National Institute of Polar Research, Tokyo, Japan Transdisciplinary Research Integration Center, Tokyo, Japan.
authors.forename: Takahiro
authors.initials: T
authors.lastname: Segawa
authors.affiliation: Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan.
authors.forename: Ayano
authors.initials: A
authors.lastname: Kobayashi
authors.affiliation: Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan.
authors.forename: Takayuki
authors.initials: T
authors.lastname: Miura
authors.affiliation: Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan.
authors.forename: Daisuke
authors.initials: D
authors.lastname: Sano
authors.affiliation: Division of Environmental Engineering, Faculty of Engineering, Hokkaido University, Sapporo, Hokkaido, Japan.
authors.forename: Satoshi
authors.initials: S
authors.lastname: Okabe
mesh_terms: D007564:Japan; D044085:Microfluidics / Q000379:methods*; D008969:Molecular Sequence Data; D010802:Phylogeny; D016133:Polymerase Chain Reaction / Q000379:methods*; D045483:Rivers / Q000821:virology*; D014780:Viruses / Q000145:classification / Q000235:genetics / Q000302:isolation & purification*; D062065:Wastewater / Q000821:virology
chemical_list: D062065:Wastewater