{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,3]],"date-time":"2025-10-03T12:51:14Z","timestamp":1759495874707,"version":"3.37.3"},"reference-count":23,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2020,11,5]],"date-time":"2020-11-05T00:00:00Z","timestamp":1604534400000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2020,11,5]],"date-time":"2020-11-05T00:00:00Z","timestamp":1604534400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"funder":[{"DOI":"10.13039\/100010661","name":"Horizon 2020 Framework Programme","doi-asserted-by":"publisher","award":["713660 PRONKJEWAIL"],"award-info":[{"award-number":["713660 PRONKJEWAIL"]}],"id":[{"id":"10.13039\/100010661","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Sci Rep"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p><jats:italic>Staphylococcus aureus<\/jats:italic> bacteraemia (SAB) is associated with high mortality and morbidity rates. Yet, there is currently no adequate diagnostic test for early and rapid diagnosis of SAB. Therefore, this study was aimed at exploring the potential for clinical implementation of a nuclease-activatable fluorescent probe for early diagnosis of SAB. To this end, clinical blood culture samples from patients with bloodstream infections were incubated for 1\u00a0h with the \u201csmart\u201d activatable P2&amp;3TT probe, the total assay time being less than 2\u00a0h. Cleavage of this probe by the secreted <jats:italic>S. aureus<\/jats:italic> enzyme micrococcal nuclease results in emission of a readily detectable fluorescence signal. Incubation of <jats:italic>S. aureus<\/jats:italic>-positive blood culture samples with the P2&amp;3TT probe resulted in 50-fold higher fluorescence intensity levels than incubation with culture-negative samples. Moreover, incubation of the probe with non-<jats:italic>S. aureus<\/jats:italic>-positive blood cultures yielded essentially background fluorescence intensity levels for cultures with Gram-negative bacteria, and only\u2009~\u20093.5-fold increased fluorescence intensity levels over background for cultures with non-<jats:italic>S. aureus<\/jats:italic> Gram-positive bacteria. Importantly, the measured fluorescence intensities were dose-dependent, and a positive signal was clearly detectable for <jats:italic>S. aureus<\/jats:italic>-positive blood cultures with bacterial loads as low as\u2009~\u20097,000 colony-forming units\/mL. Thus, the nuclease-activatable P2&amp;3TT probe distinguishes clinical <jats:italic>S. aureus<\/jats:italic>-positive blood cultures from non-<jats:italic>S. aureus<\/jats:italic>-positive blood cultures and culture-negative blood, accurately, rapidly and with high sensitivity. We conclude that this probe may enhance the diagnosis of SAB.<\/jats:p>","DOI":"10.1038\/s41598-020-76254-4","type":"journal-article","created":{"date-parts":[[2020,11,5]],"date-time":"2020-11-05T11:04:33Z","timestamp":1604574273000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":5,"title":["The smart activatable P2&amp;3TT probe allows accurate, fast, and highly sensitive detection of Staphylococcus aureus in clinical blood culture samples"],"prefix":"10.1038","volume":"10","author":[{"given":"Marina","family":"L\u00f3pez-\u00c1lvarez","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marjolein","family":"Heuker","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jorrit W. A.","family":"Schoenmakers","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Gooitzen M.","family":"van Dam","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"suffix":"II","given":"James O.","family":"McNamara","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Jan Maarten","family":"van Dijl","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marleen","family":"van Oosten","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"297","published-online":{"date-parts":[[2020,11,5]]},"reference":[{"key":"76254_CR1","doi-asserted-by":"publisher","first-page":"944","DOI":"10.1016\/j.cmi.2018.05.007","volume":"24","author":"N Peker","year":"2018","unstructured":"Peker, N., Couto, N., Sinha, B. & Rossen, J. W. Diagnosis of bloodstream infections from positive blood cultures and directly from blood samples: recent developments in molecular approaches. Clin. Microbiol. Infect. 24, 944\u2013955 (2018).","journal-title":"Clin. Microbiol. Infect."},{"key":"76254_CR2","doi-asserted-by":"publisher","first-page":"646","DOI":"10.1016\/j.arcmed.2005.02.005","volume":"36","author":"GML Bearman","year":"2005","unstructured":"Bearman, G. M. L. & Wenzel, R. P. Bacteremias: a leading cause of death. Arch. Med. Res. 36, 646\u2013659 (2005).","journal-title":"Arch. Med. Res."},{"key":"76254_CR3","doi-asserted-by":"publisher","first-page":"309","DOI":"10.1086\/421946","volume":"39","author":"JG Bartlett","year":"2004","unstructured":"Bartlett, J. G. Nosocomial bloodstream infections in US hospitals: analysis of 24,179 cases from a prospective nationwide surveillance study. Infect. Dis. Clin. Pract. 39, 309\u2013317 (2004).","journal-title":"Infect. Dis. Clin. Pract."},{"key":"76254_CR4","doi-asserted-by":"publisher","first-page":"e0157234","DOI":"10.1371\/journal.pone.0157234","volume":"11","author":"EL Burghardt","year":"2016","unstructured":"Burghardt, E. L. et al. Rapid, culture-free detection of Staphylococcus aureus Bacteremia. PLoS ONE 11, e0157234 (2016).","journal-title":"PLoS ONE"},{"key":"76254_CR5","doi-asserted-by":"publisher","first-page":"362","DOI":"10.1128\/CMR.05022-11","volume":"25","author":"SJ van Hal","year":"2012","unstructured":"van Hal, S. J. et al. Predictors of mortality in Staphylococcus aureus Bacteremia. Clin. Microbiol. Rev. 25, 362\u2013386 (2012).","journal-title":"Clin. Microbiol. Rev."},{"issue":"948","key":"76254_CR6","first-page":"e1","volume":"22","author":"P Braquet","year":"2016","unstructured":"Braquet, P. et al. Factors associated with 12 week case-fatality in Staphylococcus aureus Bacteraemia: a prospective cohort study. Clin. Microbiol. Infect. 22(948), e1-948.e7 (2016).","journal-title":"Clin. Microbiol. Infect."},{"key":"76254_CR7","doi-asserted-by":"publisher","first-page":"269","DOI":"10.1128\/CMR.3.3.269","volume":"3","author":"P Yagupsky","year":"1990","unstructured":"Yagupsky, P. & Nolte, F. S. Quantitative aspects of septicemia. Clin. Microbiol. Rev. 3, 269\u2013279 (1990).","journal-title":"Clin. Microbiol. Rev."},{"key":"76254_CR8","doi-asserted-by":"publisher","first-page":"595","DOI":"10.1111\/j.1699-0463.1993.tb00152.x","volume":"101","author":"B Jonsson","year":"1993","unstructured":"Jonsson, B., Nyberg, A. & Henning, C. Theoretical aspects of detection of bacteraemia as a function of the volume of blood cultured. APMIS 101, 595\u2013601 (1993).","journal-title":"APMIS"},{"key":"76254_CR9","doi-asserted-by":"publisher","first-page":"864","DOI":"10.1128\/JCM.17.5.864-869.1983","volume":"17","author":"NK Henry","year":"1983","unstructured":"Henry, N. K. et al. Microbiological and clinical evaluation of the isolator Lysis-centrifugation blood culture tube. J. Clin. Microbiol. 17, 864\u2013869 (1983).","journal-title":"J. Clin. Microbiol."},{"key":"76254_CR10","doi-asserted-by":"publisher","first-page":"1634","DOI":"10.1002\/pmic.200900313","volume":"10","author":"AK Ziebandt","year":"2010","unstructured":"Ziebandt, A. K. et al. Proteomics uncovers extreme heterogeneity in the Staphylococcus aureus exoproteome due to genomic plasticity and variant gene regulation. Proteomics 10, 1634\u20131644 (2010).","journal-title":"Proteomics"},{"key":"76254_CR11","doi-asserted-by":"publisher","first-page":"755","DOI":"10.1128\/MMBR.00008-06","volume":"70","author":"MJJB Sibbald","year":"2006","unstructured":"Sibbald, M. J. J. B. et al. Mapping the pathways to Staphylococcal pathogenesis by comparative secretomics. Microbiol. Mol. Biol. Rev. 70, 755\u2013788 (2006).","journal-title":"Microbiol. Mol. Biol. Rev."},{"key":"76254_CR12","doi-asserted-by":"publisher","first-page":"301","DOI":"10.1038\/nm.3460","volume":"20","author":"FJ Hernandez","year":"2014","unstructured":"Hernandez, F. J. et al. Noninvasive imaging of Staphylococcus aureus infections with a nuclease-activated probe. Nat. Med. 20, 301\u2013306 (2014).","journal-title":"Nat. Med."},{"key":"76254_CR13","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1038\/s41598-018-19289-y","volume":"8","author":"CWK Rosman","year":"2018","unstructured":"Rosman, C. W. K. et al. Ex vivo tracer efficacy in optical imaging of Staphylococcus aureus nuclease activity. Sci. Rep. 8, 1\u20138 (2018).","journal-title":"Sci. Rep."},{"key":"76254_CR14","doi-asserted-by":"publisher","first-page":"219","DOI":"10.1111\/j.1699-0463.1995.tb01098.x","volume":"103","author":"OG Brakstad","year":"1995","unstructured":"Brakstad, O. G., M\u00e6Land, J. A. & Chesneau, O. Comparison of tests designed to identify Staphylococcus aureus thermostable nuclease. APMIS 103, 219\u2013224 (1995).","journal-title":"APMIS"},{"key":"76254_CR15","doi-asserted-by":"crossref","first-page":"1016","DOI":"10.1016\/S0021-9258(18)96225-3","volume":"242","author":"JN Heins","year":"1967","unstructured":"Heins, J. N., Suriano, J. R., Taniuchi, H. & Anfinsen, C. B. Characterization of a nuclease produced by Staphylococcus aureus. J. Biol. Chem. 242, 1016\u20131022 (1967).","journal-title":"J. Biol. Chem."},{"key":"76254_CR16","doi-asserted-by":"publisher","first-page":"174","DOI":"10.1016\/j.micres.2012.09.003","volume":"168","author":"Y Hu","year":"2013","unstructured":"Hu, Y. et al. Characterization and comparative analysis of a second thermonuclease from Staphylococcus aureus. Microbiol. Res. 168, 174\u2013182 (2013).","journal-title":"Microbiol. Res."},{"key":"76254_CR17","doi-asserted-by":"publisher","first-page":"e95574","DOI":"10.1371\/journal.pone.0095574","volume":"9","author":"MR Kiedrowski","year":"2014","unstructured":"Kiedrowski, M. R. et al. Staphylococcus aureus Nuc2 is a functional, surface-attached extracellular nuclease. PLoS ONE 9, e95574 (2014).","journal-title":"PLoS ONE"},{"key":"76254_CR18","doi-asserted-by":"publisher","first-page":"1305","DOI":"10.1038\/s41598-018-19289-y","volume":"8","author":"CWK Rosman","year":"2018","unstructured":"Rosman, C. W. K. et al. Ex vivo tracer efficacy in optical imaging of Staphylococcus aureus nuclease activity. Sci. Rep. 8, 1305 (2018).","journal-title":"Sci. Rep."},{"key":"76254_CR19","first-page":"1173","volume":"1","author":"EM Trecarichi","year":"2012","unstructured":"Trecarichi, E. M., Cauda, R. & Tumbarello, M. Bloodstream Infect. 1, 1173\u20131189 (2012).","journal-title":"Bloodstream Infect."},{"key":"76254_CR20","doi-asserted-by":"publisher","first-page":"245","DOI":"10.1016\/j.jinf.2003.10.010","volume":"48","author":"O Lesens","year":"2004","unstructured":"Lesens, O. et al. Positive surveillance blood culture is a predictive factor for secondary metastatic infection in patients with Staphylococcus aureus Bacteraemia. J. Infect. 48, 245\u2013252 (2004).","journal-title":"J. Infect."},{"key":"76254_CR21","doi-asserted-by":"publisher","first-page":"41","DOI":"10.1016\/0378-1119(94)90320-4","volume":"145","author":"O Chesneau","year":"1994","unstructured":"Chesneau, O. & Solh, N. E. Primary structure and biological features of a thermostable nuclease isolated from Staphylococcus hyicus. Gene 145, 41\u201347 (1994).","journal-title":"Gene"},{"key":"76254_CR22","doi-asserted-by":"publisher","first-page":"5232","DOI":"10.1093\/nar\/20.19.5232","volume":"20","author":"O Chesneau","year":"1992","unstructured":"Chesneau, O. & Solh, N. E. Nucleotide sequence of a Nuc gene encoding the thermonuclease of Staphylococcus intermedius. Nucl. Acids Res. 20, 5232 (1992).","journal-title":"Nucl. Acids Res."},{"key":"76254_CR23","doi-asserted-by":"publisher","first-page":"4036","DOI":"10.1128\/JCM.02327-14","volume":"52","author":"F Schaumburg","year":"2014","unstructured":"Schaumburg, F. et al. Characterization of a novel thermostable nuclease homolog (NucM) in a highly divergent Staphylococcus aureus Clade. J. Clin. Microbiol. 52, 4036\u20134038 (2014).","journal-title":"J. Clin. Microbiol."}],"container-title":["Scientific Reports"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-76254-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-76254-4","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-76254-4.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2022,12,6]],"date-time":"2022-12-06T22:25:19Z","timestamp":1670365519000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s41598-020-76254-4"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,11,5]]},"references-count":23,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2020,12]]}},"alternative-id":["76254"],"URL":"https:\/\/doi.org\/10.1038\/s41598-020-76254-4","relation":{},"ISSN":["2045-2322"],"issn-type":[{"type":"electronic","value":"2045-2322"}],"subject":[],"published":{"date-parts":[[2020,11,5]]},"assertion":[{"value":"15 March 2020","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"25 September 2020","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"5 November 2020","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"J.O.M. is founder and CEO of Nuclease Probe Technologies Inc., and he holds patents on oligonucleotide-based nuclease-activatable probes for detection of bacterial nucleases. G.M.v.D. is CEO, founder and shareholder of AxelaRx\/TRACER group, CSO AxelaRx Biosciences Inc. The other authors have no conflicts of interest to disclose.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"19216"}}