Handheld FRET-Aptamer Sensor for Water Safety

Operational Technologies Corporation (OpTech) proposes to expand its current NASA Phase 2 SBIR handheld fluorometer and bone marker fluorescence resonance energy transfer (FRET)-DNA aptamer assay system to include detection of bacteria, fungi, and parasites that may contaminate astronauts' water supplies. For Gram positive bacteria, teichoic acids and peptidoglycan will serve as targets. For Gram negative bacteria, common lipopolysaccharide moieties such as 2-keto-3-deoxyoctanate (KDO antigen) will be targeted for aptamer development. Similarly, for fungi, cell wall chitin will be used to select highly specific FRET-aptamers from a randomized DNA library. Parasites such as Cryptosporidium and Giardia will require more specific whole cell or surface protein aptamer selection approaches, but OpTech has recently demonstrated detection of 30 E. coli bacteria per ml using such an approach under NSF Phase 1 SBIR funding. Prototype assays will be lyophilized in plastic cuvettes and capped under vacuum or otherwise sealed to prevent opening in negative pressure environments. Lyophilization with trehalose or other excipients will extend shelf-life to greater than 2 years for these rapid (15 minute) one step (homogeneous) FRET assays that will be quantified with an ultrasensitive commercial handheld fluorometer. Data can be displayed on the handheld reader and downloaded to a laptop computer.

OpTech is already successfully working on portable FRET-aptamer technology to rapidly (less than 15 minutes) detect fecal bacteria in water supplies and wishes to expand its repertoire of assays to include parasite and general fungal detection. The FRET-aptamer and handheld sensor package has many other applications in clinical point-of-care diagnostics, portable veterinary diagnostics, environmental pollution and chem-bio terrorism or military detection. In addition, aptamers that bind lipopolysaccharides (endotoxin) and other bacterial components could be very useful in antibacterial or antibiotic-like therapies against life-threatening conditions like sepsis and antibiotic-resistant infections.

In addition, to its present intended use for monitoring bone loss in astronauts under OpTech's Phase 2 SBIR funding from NASA, this same platform technology could be used as a portable means to monitor water quality by ultrasensitive detection of bacteria, fungi and specific parasites. Clearly also, NASA could monitor other non-bone-related clinical analytes in urine or serum for astronauts during spaceflight with one step push-button ease, if appropriate FRET-assays were developed.

This is early/mid-stage (TRL 6) — the most realistic path in is NASA SBIR/STTR, which funds small businesses and research institutions to develop technology aligned with NASA's needs (equity-free, phased funding). Check whether a current SBIR/STTR solicitation topic overlaps with this project's technology area, or contact the project directly (above) to ask.

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