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

2025-05-10Artigo científico Acesso aberto
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datacite.subject.fosCiências Naturais
dc.contributor.authorGómez, Livia
dc.contributor.authorBrandão, João
dc.contributor.authorNavarro, Anna
dc.contributor.authorMartines, Antonino
dc.contributor.authorSanseverino, Isabella
dc.contributor.authorPorcel-Rodríguez, Elena
dc.contributor.authorMarinov, Dimitar
dc.contributor.authorAbleitner, Oksana
dc.contributor.authorAllerberger, Franz
dc.contributor.authorAtsma, Adrie
dc.contributor.authorBriancesco, Rossella
dc.contributor.authorCenov, Arijana
dc.contributor.authorCíchová, Marianna
dc.contributor.authorCoccia, Anna Maria
dc.contributor.authorLibera, Simonetta Della
dc.contributor.authorDe Maeyer, Katrien
dc.contributor.authorDe Vries, Hendrik Jan
dc.contributor.authorElsinga, Goffe
dc.contributor.authorHeijnen, Leo
dc.contributor.authorHokajärvi, Anna-Maria
dc.contributor.authorJanská, Veronika
dc.contributor.authorHrabar, Jerko
dc.contributor.authorHuse-Kutowska, Monika
dc.contributor.authorJozic, Slaven
dc.contributor.authorKlatovsky, Stefanie
dc.contributor.authorKornschober, Christian
dc.contributor.authorKohls, Katharina
dc.contributor.authorKrom, Adrie
dc.contributor.authorLuden, Katrin
dc.contributor.authorMikula-Pratschke, Claudia
dc.contributor.authorPitkänen, Tarja
dc.contributor.authorPoje, Mateja
dc.contributor.authorProkšová, Miloslava
dc.contributor.authorRetelj, Matjaz
dc.contributor.authorRozwadowska, Beata
dc.contributor.authorRytkönen, Annastiina
dc.contributor.authorSarekoski, Anniina
dc.contributor.authorSchlager, Sabine
dc.contributor.authorSpringer, Burkhard
dc.contributor.authorStojan, Iva
dc.contributor.authorStroobach, Eline
dc.contributor.authorŠućić, Neven
dc.contributor.authorValério, Elisabete
dc.contributor.authorVan Pelt, Gabi
dc.contributor.authorVranken, Jerom
dc.contributor.authorVrdoljak Tomaš, Ana
dc.contributor.authorVukić Lušić, Darija
dc.contributor.authorWłos, Ewa
dc.contributor.authorWubbels, Gerhard
dc.contributor.authorZiombska, Maria
dc.contributor.authorZelenik, Katja
dc.contributor.authorZiuziakowski, Marcin
dc.contributor.authorLettieri, Teresa
dc.date.accessioned2026-02-03T15:37:56Z
dc.date.available2026-02-03T15:37:56Z
dc.date.issued2025-05-10
dc.description.abstractThe 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.eng
dc.description.abstractHighlights: -Real-time RT-PCR assay enables E. coli detection in drinking water within 6 h. -Two interlaboratory studies were performed at EU-representative level. -The real-time RT-PCR assay showed a sensitivity higher than 91 %. -RNA extraction is the most critical step of the assay. -Training and practical experience are fundamental to reach the highest sensitivity.eng
dc.description.sponsorshipThis study has been funded by the Institutional Research program of the European Commission Joint Research Centre (JRC).
dc.identifier.citationEnviron Res. 2025 Aug 15;279(Pt 2):121786. doi: 10.1016/j.envres.2025.121786. Epub 2025 May 10
dc.identifier.doi10.1016/j.envres.2025.121786
dc.identifier.issn0013-9351
dc.identifier.pmid40355062
dc.identifier.urihttp://hdl.handle.net/10400.18/10799
dc.language.isoeng
dc.peerreviewedyes
dc.publisherElsevier
dc.relation.hasversionhttps://www.sciencedirect.com/science/article/pii/S0013935125010370?via%3Dihub
dc.rights.urihttp://creativecommons.org/licenses/by-nc-nd/4.0/
dc.subjectAlternative Method Validation
dc.subjectDetection
dc.subjectDrinking Water
dc.subjectEscherichia Coli
dc.subjectFaecal Contamination
dc.subjectReal-Time Reverse Transcription PCR
dc.subjectÁgua e Solo
dc.subjectAvaliação do Risco
dc.titleApplication of a real-time reverse transcription polymerase chain reaction for rapid detection of Escherichia coli in drinking water: an EU representative studyeng
dc.typejournal article
dcterms.referenceshttps://ars.els-cdn.com/content/image/1-s2.0-S0013935125010370-mmc1.docx
dspace.entity.typePublication
oaire.citation.issuePart 2
oaire.citation.startPage121786
oaire.citation.titleEnvironmental Research
oaire.citation.volume279
oaire.versionhttp://purl.org/coar/version/c_970fb48d4fbd8a85

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