Application of a real-time reverse transcription polymerase chain reaction for rapid detection of Escherichia coli in drinking water: an EU representative study
Fast detection metod for pahtogen for E. coli in drinking water
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.
GOMEZ CORTES Livia;
BRANDÃO João;
NAVARRO Anna;
MARTINES Antonino;
SANSEVERINO Isabella;
PORCEL RODRIGUEZ Elena;
MARINOV Dimitar;
ABLEITNER Oksana;
ALLERBERGER Franz;
ATSMA Adrie;
BRIANCESCO Rossella;
CENOV Arijana;
CÍCHOVÁ Marianna;
COCCIA Anna Maria;
DELLA LIBERA Simonetta;
DE MAEYER Katrien;
DE VRIES Hendrik Jan;
ELSINGA Goffe;
HOKAJÄRVI Anna-Maria;
JANSKÁ Veronika;
HRABAR Jerko;
HUSE-KUTOWSKA Monika;
SLAVEN Josić;
KLATOVSKYAND Stefanie;
KORNSCHOBER Christian;
KOHLS Katharina;
KROM Adrie;
LUDEN Katrin;
MIKULA-PRATSCHKE Claudia;
PITKÄNEN Taria;
POJE Mateja;
PROKŠOVÁ Miloslava;
RETELJ Matjaz;
ROZWADOWSKA Beata;
RYTKÖNEN Annastiina;
SAREKOSKI Anniina;
SCHLAGER Sabine;
SPRINGER Burkhard;
STOJAN Iva;
STROOBACH Eline;
SUČIĆ Neven;
VALÉRIO Elisabete;
VAN PELT Gabi;
VRANKEN Jerom;
VRDOLJAK TOMAŠ Ana;
DARIJA VUKIĆ LUŠIĆ Darija;
WŁOS Ewa;
WUBBELS Gerhard;
ZIOMBSKA Maria;
ZELENIK Katja;
ZIUZIAKOWSKI Marcin;
LETTIERI Teresa;
2025-12-04
ACADEMIC PRESS INC ELSEVIER SCIENCE
JRC139333
1096-0953 (online),
https://www.sciencedirect.com/science/article/pii/S0013935125010370,
https://publications.jrc.ec.europa.eu/repository/handle/JRC139333,
10.1016/j.envres.2025.121786 (online),
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