The rapid detection of weaponized bacteria and toxins is a major problem during a biological attack. Although sensitive detection formats exist for many biowarfare agents, they often require advanced training and complex procedures. Luna has developed simple, rapid means for determining the presence of pathogens and bacterial toxins in water supplies using fluorescence-based assays that can be adapted for field use. The batteries of rapid assays are designed for i)
determining cell viability and bacterial loads by exploiting metabolic markers (e.g., acid-production, redox potentials,
etc) and ii) detecting bacterial toxins using fluorescent, polymerized affinity liposomes (fluorosomes). The viability
assays were characterized using E. coli, S. aureus and the anthrax simulant, B. globigii. The viability assays detected bacterial loads of ~ 10<sup>4</sup> CFU/ml and with simple filtration ~ 100CFU/ml could be detected. The affinity fluorosomes were characterized using cholera toxin (CT). Affinity liposomes displaying GM1 and anti-CT antibodies could detect CT at <μg/ml levels. Stability studies showed that affinity vesicles could be stored for weeks at 4°C or freeze-dried with no significant loss of binding capacity. Using an in-house fiber optic fluorescence system, Luna characterized the binding
of affinity fluorosomes to respective targets and determined the responses of bacterial loads in the fluorescent viability assays. Using this two-tiered approach, Luna demonstrated that water susceptible to sabotage could be easily monitored and confirmed for specific agents using simple, general and specific fluorescence-based detection schemes based on metabolism and ligand-target interactions.