Fast, sensitive, and reliable detection of waterborne pathogens by digital PCR after coagulation and foam concentration

Atsushi Jikumaru, Satoshi Ishii, Tomoko Fukudome, Yasuhiko Kawahara, Atsushi Iguchi, Yoshifumi Masago, Kei Nukazawa, Yoshihiro Suzuki

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

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 languageEnglish (US)
Pages (from-to)76-81
Number of pages6
JournalJournal of Bioscience and Bioengineering
Volume130
Issue number1
DOIs
StatePublished - 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
  • Fresh Water/chemistry
  • Sensitivity and Specificity
  • Shiga-Toxigenic Escherichia coli/genetics
  • Shiga Toxin/genetics
  • Polymerase Chain Reaction/instrumentation

PubMed: MeSH publication types

  • Journal Article
  • Evaluation Study

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