The CYTO-WATER platform integrates an automatic water concentration system that includes Celltrap® concentrating filters, a microfluidic system for automatic sample preparation and a fluorescence image cytometer.
Before the integration, the different modules have been adapted and validated individually in the laboratory to make sure that they meet the market specifications and comparing with conventional analysis and alternative methods, to determine whether the methods are suitable for detecting and quantifying microorganisms in water samples.
Regarding the concentrator module, the objective was to adapt the existing automated liquid concentration system to the needs of the CYTO-WATER platform. The bacteria contained in the water volume to be analyzed (100 mL for E. coli and 1 L for Legionella) are concentrated in a volume from 1000 to 1500μL with a recovery rate of around 50%.
For sample preparation, a microfluidic module was designed and validated. The microfluidic cartridge and tubing material was evaluated basing on the study of the optical properties, mechanical properties, hydrophilic/hydrophobic properties, and biocompatibility.
The microfluidic module was evaluated comparing samples labelled with the microfluidic system and samples labelled manually. Finally, a semi-automated as well as a fully automated, one channel microfluidic labelling systems were designed and manufactured.
The reader was adapted to the marker requirements carrying out several adjustments and improvements of hardware and software in the detection module. These adjustments were required to achieve the required performance in terms of limit of detection (LOD), sensitivity, repeatability, reproducibility and robustness, for the validation process.
The detector was validated with spiked samples to verify the main parameters of the system. The reader was also validated with natural samples, as cooling water and seawater samples. The obtained parameters in the validation tests indicated good correlations (R2>0.9) with traditional methods (culture isolation) and alternative methods, as flow cytometry (FCM), and its suitability for detection and quantification of Legionella and E. coli in natural samples.
The reader, the water concentration module and the microfluidic device (hardware and software) were integrated in an outdoor rack cabinet resulting in the CYTO-WATER platform.
The operational functioning and performance of the CYTO-WATER platform in real conditions was evaluated. A detailed laboratory and on-site validations in different facilities was performed. The validation was performed comparing results with results obtained by other methodologies (culture isolation as the standard method, and FCM and qPCR as alternative methods).
The three modules as separate devices are also very interest in terms of market opportunities. The main advantage of this approach is that each of the modules can be adapted to different technologies and can target other markets. Hence, this approach is more flexible and with a wider market in some cases. One clear example is the automatic labelling module that can be adapted to microorganisms detection with other techniques (different form the CYTO-WATER reader) and then adapted to the workflow in laboratories of different sectors, hospitals, universities, etc.
An environmental and economic comparison between the traditional spot sampling method and the CYTO-WATER system was assessed through to a Life Cycle Assessment (LCA) and Life Cycle Costing (LCC). This comparison was undertaken for the detection of Legionella in cooling towers and hot water distribution systems from hospitals and E. coli in bathing waters. Finally, results were summarized and evaluated in the validation document.
Exploitation and dissemination activities: Project brochure; 4 electronic newsletters; Roll-up; website; project video; Social media (Twitter); participation in conferences and workshops; CYTO-WATER Final Workshop was held in Barcelona; and meetings and interviews with different end-users
