Application of a real-time reverse transcription polymerase chain reaction for rapid detection of Escherichia coli in drinking water: an EU representative study

The microbiological quality of water intended for human consumption in the EU is regulated by the recast Drinking Water Directive 2020/2184 (DWD), which sets strict parametric values for intestinal enterococci and Escherichia coli (E. coli), allowing no more than zero colony-forming units per 100 mL. Detection and enumeration of E. coli typically rely on culture-based reference methods or the most probable number approach, which require 1-2 days to produce results-potentially delaying timely action during contamination events. In contrast, molecular techniques can deliver results within hours. The DWD permits the use of alternative methods if they are as reliable as the reference method and developing and validating such methods requires multiple laboratory trials in line with ISO standard 16140-2. Following this, we conducted a representative EU study to validate a molecular method based on real-time reverse transcription polymerase chain reaction for rapid E. coli detection in drinking water. In a concerted action, the first of its kind for drinking water, nineteen laboratories across ten Member States participated. To ensure consistency, drinking water was artificially contaminated with E. coli. The alternative method showed slightly lower sensitivity than the reference method (91.1 % vs. 97.2 %) but delivered much faster results, making it a valuable screening tool. It can support rapid decision-making during contamination events, reducing the risk of waterborne outbreaks and protecting public health. For reliable routine performance, appropriate training in this alternative method is strongly recommended.

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Publication Details

Journal
Environmental Research
Published
2025-05-10
DOI
https://doi.org/10.1016/j.envres.2025.121786
Primary Topic
Fecal contamination and water quality
Type
article
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article

Application of a real-time reverse transcription polymerase chain reaction for rapid detection of Escherichia coli in drinking water: an EU representative study

Burkhard Springer, Anna-Maria Hokajärvi, Marianna Cíchová, Elisabete Valério et al.
Environmental Research
Fecal contamination and water quality
article

Application of a real-time reverse transcription polymerase chain reaction for rapid detection of Escherichia coli in drinking water: an EU representative study

Burkhard Springer, Anna-Maria Hokajärvi, Marianna Cíchová, Elisabete Valério, Anna Maria Coccia, Rossella Briancesco, Sabine Schlager, Slaven Jozić, Christian Kornschober, Goffe Elsinga, Katja Zelenik, Tarja Pitkänen, Katrien De Maeyer, Gabi W. van Pelt, Oksana Ableitner, Katrin Luden, Anniina Sarekoski, João Brandão, Miloslava Prokšová, Veronika Janská, Matjaž Retelj, Isabella Sanseverino, Adrie Atsma, Iva Stojan, Beata Rozwadowska, Dimitar Marinov, Darija Vukić Lušić, Leo Heijnen, Livia Gómez, Jerko Hrabar, Neven Sučić, Teresa Lettieri, S. Della Libera, Gerhard Wubbels, Ana Vrdoljak Tomaš, Claudia Mikula-Pratschke, Arijana Cenov, Franz Allerberger, Anna Navarro, Katharina Kohls, Hendrik J. de Vries, Elena Porcel-Rodríguez, Annastiina Rytkönen, Eline Stroobach, Jerom Vranken, Monika Huse-Kutowska, Stefanie Klatovsky, Adrie Krom, Mateja Poje, Antonino Martines, Ewa Włos, Maria Ziombska, Marcin Ziuziakowski
article en

Abstract

The microbiological quality of water intended for human consumption in the EU is regulated by the recast Drinking Water Directive 2020/2184 (DWD), which sets strict parametric values for intestinal enterococci and Escherichia coli (E. coli), allowing no more than zero colony-forming units per 100 mL. Detection and enumeration of E. coli typically rely on culture-based reference methods or the most probable number approach, which require 1-2 days to produce results-potentially delaying timely action during contamination events. In contrast, molecular techniques can deliver results within hours. The DWD permits the use of alternative methods if they are as reliable as the reference method and developing and validating such methods requires multiple laboratory trials in line with ISO standard 16140-2. Following this, we conducted a representative EU study to validate a molecular method based on real-time reverse transcription polymerase chain reaction for rapid E. coli detection in drinking water. In a concerted action, the first of its kind for drinking water, nineteen laboratories across ten Member States participated. To ensure consistency, drinking water was artificially contaminated with E. coli. The alternative method showed slightly lower sensitivity than the reference method (91.1 % vs. 97.2 %) but delivered much faster results, making it a valuable screening tool. It can support rapid decision-making during contamination events, reducing the risk of waterborne outbreaks and protecting public health. For reliable routine performance, appropriate training in this alternative method is strongly recommended.

Environmental ResearchVol. 279(Pt 2)
University of Lisbon (PT), European Commission (BE), Slovenian Environment Agency (SI), Medisch Centrum Leeuwarden (NL), Lower Saxony State Museum (DE), Finnish Institute for Health and Welfare (FI), Water Research Institute (SK), Niedersächsisches Landesgesundheitsamt (DE), Stichting Artsenlaboratorium en Trombosedienst (NL), Chief Inspectorate of Environmental Protection (PL), Luxembourg Institute of Science and Technology (LU), PsyQ (NL), Joint Research Centre (IT), Province of Antwerp (BE), National Institute of Health (AM), ZeClinics (Spain) (ES), National Laboratory of Health, Environment and Food (SI), KWR Water Research Institute (NL), as2con (Croatia) (HR), National Institute of Health Dr. Ricardo Jorge (PT), Institute of Biomedical Technologies (IT), Institute of Oceanography and Fisheries (HR), Water-link (Belgium) (BE), Austrian Agency for Health and Food Safety (AT)
Clean water and sanitation
Openalex Percentile: Top 89%
Fecal contamination and water quality
DOIOpen Access PDFOpenAlex
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