Microfluidic quantitative PCR for simultaneous quantification of multiple viruses in environmental water samples

Satoshi Ishii, Gaku Kitamura, Takahiro Segawa, Ayano Kobayashi, Takayuki Miura, Daisuke Sano, Satoshi Okabe

Research output: Contribution to journalArticle

47 Citations (Scopus)

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.

Original languageEnglish
Pages (from-to)7505-7511
Number of pages7
JournalApplied and environmental microbiology
Volume80
Issue number24
DOIs
Publication statusPublished - 2014 Jan 1

ASJC Scopus subject areas

  • Biotechnology
  • Food Science
  • Applied Microbiology and Biotechnology
  • Ecology

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