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A portable MicroNanoBioSystem and Instrument for ultra-fast analysis of pathogens in food: Innovation from LOVE-FOOD lab prototype to a pre-commercial instrument

Periodic Reporting for period 2 - LoveFood2Market (A portable MicroNanoBioSystem and Instrument for ultra-fast analysis of pathogens in food: Innovation from LOVE-FOOD lab prototype to a pre-commercial instrument)

Reporting period: 2017-08-01 to 2019-04-30

The development of new methodologies advancing the state of the art in foodborne pathogen detection is a challenge for scientists and technologists. Food-related disease outbreaks have a serious impact on consumers’ health and their trust to food industry. With 48 million people in the developed countries suffering annually from foodborne diseases, several of which lead to hospitalisation or even death, it is a societal priority to strengthen food-safety management by developing and implementing more efficient methodologies to prevent such cases. This project aims to meet the challenge by providing a reliable and versatile solution thanks to the convergence of micro-nano-bio systems. The LoveFood2Market (LF2M) capitalizes on several innovative concepts which have already been proven to meet the required criteria for fast, low cost and highly sensitive analysis of pathogens in food samples in a previous research project entitled LOVE-FOOD; in the latter, a credit-card size Lab-on-Chip platform was developed where all necessary steps for bacteria detection were demonstrated by using several chips working in a modular way. Specifically, bacteria capture and lysis (one chip), DNA extraction (second chip) and amplification (third chip) and finally pathogenic-DNA detection (fourth chip) were performed in less than 8 hours and without the need for skilled personnel or large, lab-based dedicated equipment.

To achieve a higher Technology Readiness Level (TRL>7) and proceed with the successful commercialisation of the LOVE-FOOD, within LF2M we proposed to further develop the LoC by integrating the bacteria lysis, DNA purification and amplification modules, as well as the biochip detection platform on a single cartridge. This system will be able to perform multi-pathogen analysis (i.e. Salmonella, Listeria, Escherichia coli and Bacillus cereus) in multiple samples and in a rapid manner. The system is meant to be developed for dairy products and meat analysis, with a strong commitment to produce a pre-industrial prototype by the end of the project. The development of such a platform would be a big step forward towards bacteria fast analysis, a process that currently requires from 13 hrs to 3 days using lab-based molecular diagnostic or standard culturing methods, respectively.
During the reporting period of the first 18 months of the project, all seven partners worked synergistically in order to carry out the related research and technology-oriented activities, produce the associated deliverables and achieve the set milestones. Equally intensive have been the efforts related to the exploitation and communication of the project results to the end-users and stakeholders. Briefly, two new systems have been developed with outstanding performance capabilities regarding the analysis time and simplicity. Both prototypes take advantage of the unique sensing properties of acoustic wave devices for the label-free detection of pathogenic DNA (Salmonella, E. coli, Listeria or B. cereus) with a sensitivity down to 1 bacterium in 25 gr of food (milk).

During the second reporting period work efforts were focused primarily on 1. the final optimisation of the parts that consisted the Lab-on-Chip, i.e. the plastic card and the docking stations, as well as the software and 2. extensive validation studies at three end-users labs. Samples tested included milk, cheese as well as formula milk. Results from the end-users were reported to the consortium, including feedback related to the process, data analysis and reliability of the method. The very rapid nature of the technology was highly commented together with the simple methodology and cost-effectiveness of the technique. Some points for further improvement were also reported, regarding the re-usibility of the plastic cards. In addition, during the last period a workshop was organised where end-users from the food-industry were invited to present industrial needs regarding foodborne pathogen detection. The first demonstration of the method was also performed at the same time. Overall, results from the project were widely disseminated to scientific and general audience through workshops, conferences, publications, one press release, exhibitions and release of one video.
The innovative Lab-on-Chip (LOC) platform and associated instrument developed within the project have been shown to possess several unique advantages comparing to standard methods: 1. improvement of the analysis time by >80%; 2. simplicity in the engineering of the LOC by employing a two-step concept, i.e. a single chip for sample pre-treatment and a sensing element for the direct detection of the DNA; 3. Simplicity in the detection assay by employing a patented acoustic approach which detects directly DNA amplicons on the device surface (no need for DNA purification or hybridization); 4. simplicity in the fabrication by using processes associated with low-energy consumption (isothermal amplification); and finally, 5. cost-effectiveness, by employing modules commercially available and simple bio-processes. Moreover, the biggest advantage of the developed product is its generic nature, opening the possibility of using it in any application related to genetic biomarkers detection, such as the diagnosis of infectious diseases and cancer biomarkers in human samples, viral detection and GMOs identification in plants and bio-safety.

The project proposes a paradigmatic change in food-analysis based on interdisciplinary and highly innovative research in order to confront the increasing urgency for faster, economical and more efficient food pathogen detection. The need for more efficient food safety management is driven by 3 main reasons: 1. a worldwide increase in outbreaks of foodborne illnesses; 2. the associated increase in healthcare costs and food-manufacturers losses; and, 3. the globalization of food supply in the developing countries introducing an emerging need for food safety testing at the location of import. Moreover, with over 270,000 companies, the food industry is one of the largest European manufacturing sectors, providing employment to over 4 million people and having an annual turnover of 956 billion euros. The proposed work is expected to have a substantial impact in strengthening the competitiveness of Europe in the food and analytical sector while alleviating the societal burden related to food poisoning.

The positive impact of the LoveFood2Market project to the European competitiveness is clearly illustrated in a recent recognition of the consortium’s work by the European Council during the meeting of the European Ministers of Science and Innovation in February of 2016. Our work was recognised and included in the list of projects that “make European citizens proud of”, by providing “ground breaking research and innovation”, “a significant and real impact on Europe’s economy and society” and contributing to “a Europe that aims to attract investors, innovative enterprises and the best researchers”. In addition, the LF2M project has been invited by the EU to participate in the Bioeconomy week on 14-17 November 2017,as part of the successful EC-projects.
Overview of LoveFood2Market process