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The bacterial load in enteral colonization may be associated with a higher probability of transmission. Here, we aimed to establish a quantitative <jats:italic>vanA\/vanB<\/jats:italic> DNA real-time PCR assay on a high-throughput system. Limits of detection (LOD), linear range and precision were determined using serial bacterial dilutions. LOD was 46.9 digital copies (dcp)\/ml for <jats:italic>vanA<\/jats:italic> and 60.8 dcp\/ml for <jats:italic>vanB<\/jats:italic>. The assay showed excellent linearity between 4.7\u2009\u00d7\u200910<jats:sup>1<\/jats:sup> and 3.5\u2009\u00d7\u200910<jats:sup>5<\/jats:sup> dcp\/ml (<jats:italic>vanA<\/jats:italic>) and 6.7\u2009\u00d7\u200910<jats:sup>2<\/jats:sup> and 6.7\u2009\u00d7\u200910<jats:sup>5<\/jats:sup> dcp\/ml (<jats:italic>vanB<\/jats:italic>). Sensitivity was 100% for <jats:italic>vanA<\/jats:italic> and <jats:italic>vanB<\/jats:italic>, with high positive predictive value (PPV) for <jats:italic>vanA<\/jats:italic> (100%), but lower\u00a0PPV for <jats:italic>vanB<\/jats:italic> (34.6%) likely due to the presence of <jats:italic>vanB<\/jats:italic> DNA positive anerobic bacteria in rectal swabs. Using the assay on enriched VRE broth <jats:italic>vanB<\/jats:italic> PPV increased to 87.2%. Quantification revealed median 2.0\u2009\u00d7\u200910<jats:sup>4<\/jats:sup> dcp\/ml in PCR positive but VRE culture negative samples and median 9.1\u2009\u00d7\u200910<jats:sup>4<\/jats:sup> dcp\/ml in VRE culture positive patients (maximum: 10<jats:sup>7<\/jats:sup> dcp\/ml). The automated <jats:italic>vanA\/B<\/jats:italic>_UTC assay can be used for <jats:italic>vanA\/vanB<\/jats:italic> detection and quantification in different diagnostic settings and may support future clinical studies assessing the impact of bacterial load on risk of infection and transmission.<\/jats:p>","DOI":"10.1038\/s41598-024-54037-5","type":"journal-article","created":{"date-parts":[[2024,2,12]],"date-time":"2024-02-12T15:04:21Z","timestamp":1707750261000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Adaptation and validation of a quantitative vanA\/vanB DNA screening assay on a high-throughput PCR system"],"prefix":"10.1038","volume":"14","author":[{"given":"Katja","family":"Giersch","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Konstantin","family":"Tanida","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Both","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dominik","family":"N\u00f6rz","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Denise","family":"Heim","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Holger","family":"Rohde","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Martin","family":"Aepfelbacher","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marc","family":"L\u00fctgehetmann","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2024,2,12]]},"reference":[{"key":"54037_CR1","doi-asserted-by":"publisher","first-page":"318","DOI":"10.1016\/S1473-3099(17)30753-3","volume":"18","author":"E Tacconelli","year":"2018","unstructured":"Tacconelli, E. et al. Discovery, research, and development of new antibiotics: The WHO priority list of antibiotic-resistant bacteria and tuberculosis. Lancet. Infect. Dis 18, 318\u2013327. https:\/\/doi.org\/10.1016\/S1473-3099(17)30753-3 (2018).","journal-title":"Lancet. Infect. Dis"},{"key":"54037_CR2","doi-asserted-by":"publisher","DOI":"10.1016\/J.DRUP.2020.100732","volume":"53","author":"G Werner","year":"2020","unstructured":"Werner, G. et al. Thirty years of VRE in Germany\u2014\u201cexpect the unexpected\u201d: The view from the National Reference Centre for Staphylococci and Enterococci. Drug Resist. Updates 53, 100732. https:\/\/doi.org\/10.1016\/J.DRUP.2020.100732 (2020).","journal-title":"Drug Resist. Updates"},{"key":"54037_CR3","doi-asserted-by":"publisher","first-page":"266","DOI":"10.1038\/NRMICRO2761","volume":"10","author":"CA Arias","year":"2012","unstructured":"Arias, C. A. & Murray, B. E. The rise of the Enterococcus: Beyond vancomycin resistance. Nat. Rev. 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D.N. received speaker honoraria and related travel expenses from Roche Diagnostics. Funding for the study was provided by Roche Molecular Systems (Pleasanton, California, USA). COBAS is a trademark of Roche. All other trademarks are the property of their respective owners. The rest of the authors does not declare any competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"3523"}}