Pîndaru Andreea, Măruțescu Luminița Gabriela, Popa Marcela, Lambert Claude, Chifiriuc Mariana-Carmen
Department of Botany and Microbiology, Faculty of Biology, University of Bucharest, 030018 Bucharest, Romania.
Research Institute of University of Bucharest, University of Bucharest, 030018 Bucharest, Romania.
Microorganisms. 2025 May 19;13(5):1156. doi: 10.3390/microorganisms13051156.
A rapid and accurate evaluation of a chemical disinfectant's 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.
快速准确地评估化学消毒剂的杀菌效果对于确保有效的感染控制、防止病原体传播以及支持新消毒剂配方的开发至关重要。在本研究中，我们报告了一种用于评估消毒剂杀菌效果的快速、无标记流式细胞术（FCM）方案。针对EN 13727+A2推荐的标准菌株和十种临床菌株，评估了五种市售消毒剂（醇类、氧化剂和烷基化剂）。这种无标记FCM方法通过评估散射光图谱（FSC-H/SSC-H）和计数差异来确定消毒剂的效果。无标记FCM在大约4小时内得出结果，并且与可能需要长达48小时的标准测试显示出很强的相关性（91.4%，灵敏度0.94，特异性0.98）。我们的结果证明了无标记FCM能够在同一天可靠地评估化学消毒剂的效果，且比当前基于生长的方法快得多。此外，该研究突出了FCM方法在检测氯基消毒剂处理后活的但不可培养细菌出现情况方面的潜力。凭借其速度、准确性以及在单细胞水平识别细菌损伤的能力，FCM方法是评估新消毒剂配方效果的有力工具。
