Phages are the natural evolved viral predators of bacteria and therefore can be useful tools to detect or kill their bacterial hosts. The phages ability for specifically recognize and infect their hosts, combined with new methods in synthetic biology and nanotechnology, has allowed the engineering of hybrid bio/inorganic nanosensors which can be used to irreversibly bind bacteria and magnetically separate them from a large sample. Additionally, we have equipped the phages with genes for sensitive reporter enzymes which they inject into their host during infection. The host-synthesized reporter enzymes are then released as the phage lyses the bacteria allowing for sensitive detection.<br/>Phages were conjugated to magnetic nanoparticles and used to separate E. coli from liquid samples. The binding of the phages to their host has evolved in broad environmental conditions and therefore, the phages demonstrated an improved separation ability compared to traditional immunomagnetic methods. The improved binding was most evident outside physiological conditions with regard to pH, salinity, and temperature.<br/>Genes for reporter enzymes which were inserted into the phages allowed for colorimetric, electrochemical, fluorescent, or luminescent detection of the bacterial host. The reporter enzymes were genetically engineered to allow for optimal expression, activity and immobilization. This has allowed detection limits of &lt; 10 CFU of bacterial hosts from 100 mL of drinking water.<br/>The ability of phages to serve as both biorecognition elements and gene delivery tools make them valuable tools in low-cost bacterial diagnostics. Future research into the engineering of host specificity will allow these tools to be used to detect and treat almost all bacterial diseases.

Development of phage-based nanobots for the recognition, separation, and detection of bacterial pathogens

255th National Meeting (2018)

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