BEACONS is a unique collaboration between the United States, Northern Ireland and the Republic of Ireland, that aims to develop a novel sample preparation device coupled to a portable sensor which will be capable of rapidly analyzing for the presence of major aquatic toxins and associated organisms in water samples. It harnesses the substantial complementary expertise of the international partners and their institutions to address a problem that has world-wide implications for human health and aquatic related industries. The results will be used for the generation of a commercially applicable prototype device that could have the potential to be applied to a wide range of different analytical problems. The program will involve significant exchange of personnel and expertise and will generate novel scientific data and approaches linked with the solution of a key problem for all the partners' jurisdictions.

This project will develop a set of easily portable instruments that can be utilized for biotoxin and toxic organism detection, and be adapted subsequently for monitoring of other potentially harmful organisms and their toxins. A key element for any field deployable sensing instrument is the acquisition and preparation of samples. Thus, the proposed research includes development of a sample preparation platform and associated protocols for concentration and delivery of targeted toxin and nucleic acid sequences to a hand-held detection device. The expected outcome is the development of a sample preparation module and assay methods for an optical detection device, based on planar waveguide technology, capable of measuring in real time the presence of both toxin producing organisms and the toxins themselves in coastal and fresh water samples. The sample preparation module will involve the integration of novel microfluidics with peptide nucleic acids (PNAs) and engineered broad affinity, highly stable capture antibodies. Resultant extracts will be introduced into the detection system, which will employ an array-based, planar waveguide technology previously incorporated into a hand-held device by a commercial partner and made available to project collaborators for development of assays for toxins and organisms. They will utilize highly specific, engineered antibodies for capturing toxin molecules and PNAs as capture probes to detect the corresponding toxigenic organisms. This technology is expected to provide new and improved means of assessing the potential risks associated with contamination of coastal and fresh water fishery resources by algal biotoxins.

The US-Ireland Partnership Program has been endorsed at the highest political levels by the three jurisdictions (U.S., Republic of Ireland, and Northern Ireland). The added value for the U.S. from the BEACONS proposal includes a combination of significant intellectual and financial contributions from Ireland. Moreover, the project will involve a well coordinated exchange of researchers between the participating organizations. In particular, the opportunities of exchange visits for young scientists employed on the project will be important from both scientific and cultural perspectives. A critical element of detecting any analyte is sample processing. Therefore, in addition to stronger international collaborations and training of young scientists, this project aims to provide a new portable and flexible platform that can be adapted for the sensor-based detection of other biotoxins and/or microbes.

Project Report

Our BEACONS project developed a strong collaboration between U.S. (University of Maine and NOAA- Charleston) and Irish partners (Queens University Belfast and Dublin City University). Through a joint effort with an industry partner (mBio), we identified a sensor technology that facilitated the rapid, sensitive, simultaneous, and cost-effective detection of target analytes on a small benchtop device. We have utilized several emerging technologies to accomplish this, including antibody engineering, surface plasmon resonance (SPR) in a field portable instrument the Spirit v3.1 manufactured by Seattle Sensor Systems, Inc., and the use of peptide nucleic acids (PNAs) as probes. Our target organisms are toxin producing Mycrocystis and Alexandrium species. Both of these groups have been show to contaminate either water or food products (respectively). The aim of this effort was to develop an assay capable of detecting multiple PSTs in a single analysis by functionalizing sensor chips with either saxitoxin (STX) or neosaxitoxin (NEO). The respective toxins were immobilized through an oligoethylene glycol linker (i.e., jeffamine) to carboxymethyldextran (CMD) coating the gold sensor surface or by direct linkage of toxin-protein conjugates (i.e., STX-OVA, NEO-OVA). Assay development has been ongoing for both mycrocystins (MC) and paralytic shellfish poisoning toxins (PSTs). PNA based species-specific probes directed the causative alga associated with PSTs, (Alexandrium sp.) have been developed for both SPR instrument Spirit and a hand-held Disc2(5) colorometric dye-based assay. Epitope specific antibodies directed toward MC and PSTs have been refined and applied to the mBio waveguide sensor platform. Results indicate that these antibodies are highly sensitive for their toxin targets on this platform although are considered sub-optimal on the SPR platform. PNA based probes compatible with the mBio platform are still under development.

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University of Maine
United States
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