Final Report Summary - IMMUNOLEGIO (Rapid biotechniques based on immunosensors for in situ detection of legionella in industrial and environmental water samples)
The legionella bacteria, responsible for the Legionnaires' disease, are a group of organisms present in natural water bodies in harmless concentrations. However, manmade water systems often provide suitable conditions for the bacteria amplification to levels causing disease outbreak. The mortality rate of such outbreaks in the EU ranges to up to 30 % of the infected population, thus the timely detection of the organisms' presence is of critical importance.
Nevertheless, the existing technologies do not allow for such an accurate detection due to frequent errors and large fluctuations inherent with cell computation. Moreover, quantitative counting on a selective culture is unable to offer real-time data which could enable immediate remedial actions.
The IMMUNOLEGIO project aimed to increase the effectiveness of legionella detection technology through the development of an alternative method with high sensitivity and short detection time. The proposed technique was applicable in both natural and manmade systems, provided a complete analysis within thirty minutes and allowed for repetitive measurements at different timing points. Moreover, it was automated and portable and had no requirements for specialised personnel or instrumentation.
The developed method achieved the following objectives:
1. increased the knowledge related to water monitoring and control and the sector efficiency in Europe;
2. improved public health and safety via the rapid risks' detection;
3. produced innovative outcomes in the area of biosensors which contributed to the creation of value-added jobs.
Moreover, the project was beneficial for numerous related industries and produced a series of recommended practices and standards for bacteria management systems.
The produced instrument was based on a magnetoresistive biosensor and on the use of magnetic particles as biological markers. The markers were characterised, functionalised with antibodies and laboratory validated in terms of their selectivity and specificity. Eight different commercially available paramagnetic beads were analysed and a coating procedure, applicable in all cases, was developed. The selected monoclonal antibodies were immobilised on the biosensor chip surface fabricated with gold, and the reproducibility and biological stability of the process were evaluated.
Furthermore, the project included the construction of an adequate biosensor. Hardware and software elements of the instrument were also developed, including electronics, mechanical parts, and the user interface and analysis software. The process control and data acquisition was feasible through the construction of a pocket computer. The developed prototype was evaluated at laboratory scale using real industrial samples and its assessment continued after the project completion.
Various initially selected approaches were modified during the proposal elaboration. The flow-through concept was replaced by a batch concept because of its associated costs, difficulties in handling dry samples and increased testing time in case of large sample volumes. The biosensor was redesigned since the commercial alternatives had a very short detection range; however, this required the redistribution of resources and caused significant delays and the need for extension of the project duration. Moreover, the applied electric model had to be redesigned since the integration of the system components resulted in parasitic capacitances. Hence, the full system validation was performed after the official project closure due to the supplementary activities' duration.
A market research was conducted, defining the necessity for affordable, easy to use, portable equipment which could obtain the results on site. Thus the designed criteria and technical specifications were selected to respond to the aforementioned market needs. Moreover, a cost analysis of both beads and antibodies was performed, along with proof of principle tests to ensure that the detection beads and the capture surface could be employed successfully within the project. In addition, various functionalisation strategies for antibodies' immobilisation were tested and optimised, and those providing the most promising results were further characterised so as to determine their specificity, stability and reproducibility.
Numerous activities were carried out for knowledge management and elaboration, such as patent search, preparations for certification, training, dissemination material development and technology watch actions. The patent search referred to three components of the device, namely the micro-inductive sensor, the immunosensor and the automated sample preparation unit. It was necessary to evaluate the novelty potential of the components and to avoid infringing any existing patents by the employed technologies. However, none of the developed solutions were innovative enough; hence, other means of results protection were considered. Moreover, all the essential health and safety requirements of the European Directives were met so as to certify the instrument according to the applied standards.
Training sessions were organised in order to communicate the developed knowledge to the involved partners. The distributed dissemination material included flyers, posters, publications, as well as a multimedia guide demonstrating the use of IMMUNOLEGIO. Finally, relevant products that were concurrently developed and presented to the market were compared to the prototype device and their differences were highlighted as part of the dissemination strategy.
Nevertheless, the existing technologies do not allow for such an accurate detection due to frequent errors and large fluctuations inherent with cell computation. Moreover, quantitative counting on a selective culture is unable to offer real-time data which could enable immediate remedial actions.
The IMMUNOLEGIO project aimed to increase the effectiveness of legionella detection technology through the development of an alternative method with high sensitivity and short detection time. The proposed technique was applicable in both natural and manmade systems, provided a complete analysis within thirty minutes and allowed for repetitive measurements at different timing points. Moreover, it was automated and portable and had no requirements for specialised personnel or instrumentation.
The developed method achieved the following objectives:
1. increased the knowledge related to water monitoring and control and the sector efficiency in Europe;
2. improved public health and safety via the rapid risks' detection;
3. produced innovative outcomes in the area of biosensors which contributed to the creation of value-added jobs.
Moreover, the project was beneficial for numerous related industries and produced a series of recommended practices and standards for bacteria management systems.
The produced instrument was based on a magnetoresistive biosensor and on the use of magnetic particles as biological markers. The markers were characterised, functionalised with antibodies and laboratory validated in terms of their selectivity and specificity. Eight different commercially available paramagnetic beads were analysed and a coating procedure, applicable in all cases, was developed. The selected monoclonal antibodies were immobilised on the biosensor chip surface fabricated with gold, and the reproducibility and biological stability of the process were evaluated.
Furthermore, the project included the construction of an adequate biosensor. Hardware and software elements of the instrument were also developed, including electronics, mechanical parts, and the user interface and analysis software. The process control and data acquisition was feasible through the construction of a pocket computer. The developed prototype was evaluated at laboratory scale using real industrial samples and its assessment continued after the project completion.
Various initially selected approaches were modified during the proposal elaboration. The flow-through concept was replaced by a batch concept because of its associated costs, difficulties in handling dry samples and increased testing time in case of large sample volumes. The biosensor was redesigned since the commercial alternatives had a very short detection range; however, this required the redistribution of resources and caused significant delays and the need for extension of the project duration. Moreover, the applied electric model had to be redesigned since the integration of the system components resulted in parasitic capacitances. Hence, the full system validation was performed after the official project closure due to the supplementary activities' duration.
A market research was conducted, defining the necessity for affordable, easy to use, portable equipment which could obtain the results on site. Thus the designed criteria and technical specifications were selected to respond to the aforementioned market needs. Moreover, a cost analysis of both beads and antibodies was performed, along with proof of principle tests to ensure that the detection beads and the capture surface could be employed successfully within the project. In addition, various functionalisation strategies for antibodies' immobilisation were tested and optimised, and those providing the most promising results were further characterised so as to determine their specificity, stability and reproducibility.
Numerous activities were carried out for knowledge management and elaboration, such as patent search, preparations for certification, training, dissemination material development and technology watch actions. The patent search referred to three components of the device, namely the micro-inductive sensor, the immunosensor and the automated sample preparation unit. It was necessary to evaluate the novelty potential of the components and to avoid infringing any existing patents by the employed technologies. However, none of the developed solutions were innovative enough; hence, other means of results protection were considered. Moreover, all the essential health and safety requirements of the European Directives were met so as to certify the instrument according to the applied standards.
Training sessions were organised in order to communicate the developed knowledge to the involved partners. The distributed dissemination material included flyers, posters, publications, as well as a multimedia guide demonstrating the use of IMMUNOLEGIO. Finally, relevant products that were concurrently developed and presented to the market were compared to the prototype device and their differences were highlighted as part of the dissemination strategy.
