Final Report Summary - ENVIRO (Low-cost paper-based microfluidic devices for the detection of waterborne pathogens)
The scientific aim of enVIRO was to develop novel low-cost paper-based microfluidic systems for the detection of waterborne pathogens. Norovirus and E. coli were the initial exemplar targets as they are the primary agents of global gastrointestinal infections caused by contaminated drinking and bathing water. Three-dimensional microfluidic paper analytical devices (µPADs) were implemented with isothermal nucleic acid amplification technology (INAAT) for sensitive quantitation of pathogens in environmental samples. This was only possible by joining the expertise in molecular biology and nanotechnology at the University of Southampton (UoS) with the pioneering excellence in paper-based microfluidics at the group of the inventor of the field, George Whitesides, at Harvard University (HU). The enVIRO project was primarily a research-driven training programme with a 24-month outgoing period spent at HU, for development of novel prototype 3D INAAT µPADs for environmental samples, and a re-integration phase at UoS in ERA, for knowledge dissemination and prototype implementation that addresses increasing global and European needs in tackling water contamination and infection issues.
In the first 12 months of the project, we developed a μPAD device performing immunoassays in the field. The device can detect proteins in whole blood of bovine origin. The deliverable, due in month 6, originally described a simple μPAD, but this has been updated to a superior, more complex, multiplex device for the in-situ diagnosis of zoonoses. This resulted to one published patent by the fellow. Another paper-based device (the “Sliding Strip”) that performs immunoassays was co-developed by the fellow, and this resulted to one publication. In addition, a working INAAT assay was designed and optimized for the same bacterial target (an exemplar pathogen that causes zoonoses). This completed a deliverable due in month 9.
As per the project deliverables due in month 24, the INAAT assay−developed on the bench−was adapted for field use and quantitative results on a 3D μPAD with a handheld, quantitative electrochemical analyser, which was also co-developed by the fellow. This deliverable was originally planned for the end of month 24 and only included semi-quantitative detection by eye. The work is novel and resulted in a second published patent for the fellow. The final deliverable for the month 24 was evaluation of the prototype by an independent stakeholder; the latter concluded evaluation of the prototype at their facilities. One of the collaborating stakeholder organizations is a named co-inventor on the patent and co-author on the resulting publications. In addition, at least four manuscripts for high-impact journals were drafted with the results of all work conducted during the first 24 months, two of which have already been published by February 2018. Considerable intellectual property was developed and four patent applications have been filed in total. The deliverable due in month 36, which was a lab demonstration of prototype 3D INAAT µPAD, was completed ahead of time. The exemplar pathogen was adapted from norovirus due to complications with real-sample availability.
Our results are impactful for field analysis of pathogens in humans and animals using both nucleic acids and proteins. Minimal infrastructure is necessary for the operation of our portable reader is self-powered. The user does not require any specialist knowledge and once the−potentially pathogenic−sample is added to the device, the device is fully sealed. All the components of the device are disposable by simple incineration. A summary of the project findings is at nefelitsaloglou.eu.
In the first 12 months of the project, we developed a μPAD device performing immunoassays in the field. The device can detect proteins in whole blood of bovine origin. The deliverable, due in month 6, originally described a simple μPAD, but this has been updated to a superior, more complex, multiplex device for the in-situ diagnosis of zoonoses. This resulted to one published patent by the fellow. Another paper-based device (the “Sliding Strip”) that performs immunoassays was co-developed by the fellow, and this resulted to one publication. In addition, a working INAAT assay was designed and optimized for the same bacterial target (an exemplar pathogen that causes zoonoses). This completed a deliverable due in month 9.
As per the project deliverables due in month 24, the INAAT assay−developed on the bench−was adapted for field use and quantitative results on a 3D μPAD with a handheld, quantitative electrochemical analyser, which was also co-developed by the fellow. This deliverable was originally planned for the end of month 24 and only included semi-quantitative detection by eye. The work is novel and resulted in a second published patent for the fellow. The final deliverable for the month 24 was evaluation of the prototype by an independent stakeholder; the latter concluded evaluation of the prototype at their facilities. One of the collaborating stakeholder organizations is a named co-inventor on the patent and co-author on the resulting publications. In addition, at least four manuscripts for high-impact journals were drafted with the results of all work conducted during the first 24 months, two of which have already been published by February 2018. Considerable intellectual property was developed and four patent applications have been filed in total. The deliverable due in month 36, which was a lab demonstration of prototype 3D INAAT µPAD, was completed ahead of time. The exemplar pathogen was adapted from norovirus due to complications with real-sample availability.
Our results are impactful for field analysis of pathogens in humans and animals using both nucleic acids and proteins. Minimal infrastructure is necessary for the operation of our portable reader is self-powered. The user does not require any specialist knowledge and once the−potentially pathogenic−sample is added to the device, the device is fully sealed. All the components of the device are disposable by simple incineration. A summary of the project findings is at nefelitsaloglou.eu.