{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,12]],"date-time":"2026-05-12T18:50:38Z","timestamp":1778611838331,"version":"3.51.4"},"reference-count":77,"publisher":"MDPI AG","issue":"5","license":[{"start":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T00:00:00Z","timestamp":1747612800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Executive Agency for Higher Education, Research, Development and Innovation Funding, Romania-UEFISCDI","award":["PN-III-P2-2.1-PED-2021-2115 (620 PED)"],"award-info":[{"award-number":["PN-III-P2-2.1-PED-2021-2115 (620 PED)"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Microorganisms"],"abstract":"<jats:p>A rapid and accurate evaluation of a chemical disinfectant\u2019s bactericidal efficacy is crucial for ensuring effective infection control, preventing the spread of pathogens, and supporting the development of new disinfectant formulations. In this study, we report a rapid, label-free flow cytometry (FCM) protocol for evaluating the bactericidal efficacy of disinfectants. Five commercial disinfectants (alcohols, oxidizing agents, and alkylating agents) were evaluated against type strains recommended by EN 13727+A2 and ten clinical strains. The label-free FCM method allowed the determination of disinfectant efficacy through assessment of scatter light profiles (FSC-H\/SSC-H) and count differences. The label-free FCM provided the results in approximately 4 h and showed strong correlation with standard tests (91.4%, sensitivity 0.94 and specificity 0.98) that can take up to 48 h. Our results represent a proof-of-principle that label-free FCM can reliably assess the efficacy of chemical disinfectants, the same day, and substantially faster than the current growth-based methods. Additionally, the study highlights the potential of the FCM method for detecting the occurrence of viable but non-culturable bacteria following treatment with chlorine-based disinfectants. With its speed, accuracy, and capability to identify bacterial injuries at a single-cell level, the FCM method is a powerful tool for assessing the efficacy of new disinfectant formulations.<\/jats:p>","DOI":"10.3390\/microorganisms13051156","type":"journal-article","created":{"date-parts":[[2025,5,19]],"date-time":"2025-05-19T03:51:03Z","timestamp":1747626663000},"page":"1156","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["Label-Free Flow Cytometry: A Powerful Tool to Rapidly and Accurately Assess the Efficacy of Chemical Disinfectants"],"prefix":"10.3390","volume":"13","author":[{"given":"Andreea","family":"P\u00eendaru","sequence":"first","affiliation":[{"name":"Department of Botany and Microbiology, Faculty of Biology, University of Bucharest, 030018 Bucharest, Romania"},{"name":"Research Institute of University of Bucharest, University of Bucharest, 030018 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-4045-079X","authenticated-orcid":false,"given":"Lumini\u021ba Gabriela","family":"M\u0103ru\u021bescu","sequence":"additional","affiliation":[{"name":"Department of Botany and Microbiology, Faculty of Biology, University of Bucharest, 030018 Bucharest, Romania"},{"name":"Research Institute of University of Bucharest, University of Bucharest, 030018 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Marcela","family":"Popa","sequence":"additional","affiliation":[{"name":"Research Institute of University of Bucharest, University of Bucharest, 030018 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Claude","family":"Lambert","sequence":"additional","affiliation":[{"name":"Neurotoxicology, Development and Bioactivity, LCOMS\/ENOSIS, Universit\u00e9 de Lorraine, 57000 Metz, France"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-6098-1857","authenticated-orcid":false,"given":"Mariana-Carmen","family":"Chifiriuc","sequence":"additional","affiliation":[{"name":"Department of Botany and Microbiology, Faculty of Biology, University of Bucharest, 030018 Bucharest, Romania"},{"name":"Research Institute of University of Bucharest, University of Bucharest, 030018 Bucharest, Romania"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2025,5,19]]},"reference":[{"key":"ref_1","unstructured":"Jiang, X., Randolph, K.C., Osborn, S.L., and Tyler, H.L. 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