Development of a rapid method based on flow cytometry for testing disinfectants efficacy against problematic nosocomial agents
Project Director: Dr. Luminița-Gabriela Măruțescu
Project code: PN-III-P2-2.1-PED-2021-2115
Contract number: PD 620
Project title: Development of a rapid method based on flow cytometry for testing disinfectants efficacy against problematic nosocomial agents
Project Acronym: FAST-DET
Project Duration: 24 months
The total buget of the project: 600,000 RON
The starting date of the contract: 21/06/2022
The date of conclusion of the contract: 21/06/2024
Abstract
Healthcare-associated infections (HAI) are among the priority public health problems al global level, by the high morbidity and mortality rates, often occurred as a consequence of resistance development, generating important economic and social burden. Thus, resistance to disinfectants and co-/cross-selection for antibiotic resistance (AR) are serious problems in managing infection control. The project addresses the limitations of standard, time-consuming, culture-based methodologies, by proposing the development and validation of a flow cytometry method (FCM) for disinfectants efficacy testing, under practical, in-use conditions. The FCM method will provide a rapid and accurate tool, allowing the quantification and differentiation of live, dead or viable, but not cultivable bacteria. By providing faster results on disinfectant efficacy against antibiotic resistant nosocomial pathogens, the FCM method will assure a more rapid and efficient selection of an efficient disinfectant, and thus contribute to the prevention of cross-contamination and spread of HAI, as well as to the reduction of the selective pressure for AR. For achieving the above aim, we propose the following specific objectives: O1. Design and optimization of the FCM experimental protocol for disinfectant efficacy testing. O2. Validation of the FCM method against the current CEN standard methods. We will investigate if the damaged cells resuscitate and grow in some environmental conditions or at least retain their living properties and remain non-culturable. Highlighting the presence of such microbial populations on a surface could provide a very effective tool for investigating, controlling and limiting the environmental charge of bacteria with potential to spread and cause nosocomial infections, allowing more rapid and efficient response actions (saving time, money and/or reducing number of deaths).
Project Objective
The project aims at addressing the limitations associated with traditional, culture-based methodologies for disinfectants efficacy testing by proposing the development and validation of a clinical flow cytometry method for rapid and accurate quantification of live and dead bacteria for disinfectant efficacy testing, under practical, in-use conditions. The flow cytometry method (FCM) for disinfectant efficacy can be performed in just a few hours, with high accuracy. By providing faster results on disinfectant efficacy against antibiotic resistant nosocomial pathogens, the FCM method will assure a more rapid and efficient response for the selection of an efficient disinfectant, and thus contribute to the prevention of cross-contamination and spread of HAIs and reduction of the pressure for antibiotic resistance.
For achieving the above aim we need to reach following specific objectives:
O1. Design and optimization of the FCM experimental protocol for disinfectant efficacy testing.
O2. Validation of the FCM method against the current CEN standard methods.
Expected results
The project FAST-DET will provide a rapid and accurate method for the assessment of disinfectants’ efficiency against nosocomial pathogens. Furthermore, FCM method could provide insights into damaged bacterial cell populations. We will investigate if the damaged cells resuscitate and grow in some environmental conditions or at least retain their living properties and remain non-culturable. In this way, by highlighting the presence of such populations on a surface could provide a very effective tool for investigating, controlling and limiting the environmental biocharge of bacteria with potential to spread and cause nosocomial infections, allowing more rapid and efficient response actions (saving time and/or reducing number of death). The obtained results will make the subject of a PhD thesis; data will also be described in scientific papers (at least 4 ISI articles), a patent request (publication will be possible only after the protection of intellectual property).
The working principle of the CEN standard methods and the FCM method for disinfectant efficacy testing.
Summary_PED620_Stage1
Summary_PED620_Stage2
Scientific and technical report – Summary – Stage III
