Adapted method for rapid detection and quantification of pathogen Campylobacter jejuni from environmental water samples

Angela Sun, Paul M. Mirzayans, Andrew M. Piggott, Jo-Ann L. Stanton, Anwar Sunna*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

1 Citation (Scopus)

Abstract

Building on a previously developed workflow for rapid and sensitive pathogen detection by qPCR, this work has established a sample treatment strategy that produces consistent quantification efficiencies (QEs) for Campylobacter jejuni against a complex and highly variable sample matrix from a suburban river. The individual treatments most effective at minimizing the inhibitory effects of the sample matrix were pH buffering with HEPES (50 mM, pH 5.7) and addition of the surfactant Tween 20 (2% v/v). Unexpectedly, sample acidification (pH 4-5) resulting from the use of aged Tween 20 that had undergone partial hydrolysis, appeared to play a key role in enhancing QE. This effect could be replicated by direct pH adjustment with dilute hydrochloric acid and may be linked to the solubilization and removal of inhibitory particles at an acidic pH. While the effectiveness of each individual treatment method varied, a combined treatment of either HEPES buffer + Tween 20, or direct pH adjustment + Tween 20, consistently produced QEs of 60%-70% and up to 100%, respectively, over a sampling period of one year. The consistency and scalability of this workflow make it a suitable alternative to culture-based ISO methods for detecting Campylobacter spp.

Original languageEnglish
Article numberfiad058
Pages (from-to)1-9
Number of pages9
JournalFEMS Microbiology Ecology
Volume99
Issue number7
Early online date27 May 2023
DOIs
Publication statusPublished - Jul 2023

Keywords

  • Campylobacter jejuni
  • environmental water
  • filtration
  • qPCR
  • water testing
  • waterborne pathogen

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