Abstract
The quantification of pathogens is important for assessing water safety and preventing disease outbreaks. Culture-independent approaches, such as quantitative PCR (qPCR) and digital PCR (dPCR), are useful techniques for quantifying pathogens in water samples. However, since pathogens are usually present at low concentrations in water, it is necessary to concentrate microbial cells before extracting their DNA. Many existing microbial concentration methods are inefficient or take a long time to perform. In this study, we applied a coagulation and foam separation method to concentrate environmental water samples of between 1000 and 5000 mL to 100 μL of DNA (i.e., a 1–5 × 104-fold concentration). The concentration process took <1 h. The DNA samples were then used to quantify various target pathogens using dPCR. One gene, the Shiga toxin gene (stx2) of Shiga toxin-producing Escherichia coli, was detected at 32 copies/100 mL in a river water sample. The coagulation and foam concentration method followed by dPCR reported herein is a fast, sensitive, and reliable method to quantify pathogen genes in environmental water samples.
Original language | English (US) |
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Pages (from-to) | 76-81 |
Number of pages | 6 |
Journal | Journal of Bioscience and Bioengineering |
Volume | 130 |
Issue number | 1 |
DOIs | |
State | Published - Jul 2020 |
Bibliographical note
Funding Information:This work was supported by a Japan Science and Technology Agency (JST)-Mirai Program (Grant No. JPMJMI18DC), Japan.
Funding Information:
This work was supported by a Japan Science and Technology Agency (JST)-Mirai Program (Grant No. JPMJMI18DC ), Japan.
Publisher Copyright:
© 2020 The Society for Biotechnology, Japan
Keywords
- Coagulation and foam concentration
- Digital PCR
- Disease outbreaks
- Pathogen detection
- Shiga-toxigenic Escherichia coli
- Water quality