Pathogen-Imprinted Organosiloxane Polymers as Selective Biosensors for the Detection of Targeted E. coli.

Abstract

Early detection of pathogens requires methods that are fast, selective, sensitive and affordable. We report the development of a biosensor with high sensitivity and selectivity based on the low-cost preparation of organosiloxane (OSX) polymers imprinted with E. coli-GFP (green fluorescent protein). OSX polymers with high optical transparency, no cracking, and no shrinkage were prepared by varying several parameters of the sol-gel reaction. The unique shape and chemical fingerprint of the targeted inactivated E. coli-GFP were imprinted into bulk polymers by replication imprinting where the polymer solution was dropcast onto a bacteria template that produced a replica of the bacterial shape and chemistry on the polymer surface upon removal of the template. Capture performances were studied under non-laminar flow conditions with samples containing inactivated E. coli-GFP and compared to inactivated S. typhimurium-GFP. Capture selectivity ratios are dependent on the type of alkoxysilanes used, the H₂O:silane molar ratio, and the polymerization temperature. The bacteria concentration in suspension ranged from ~6 × 10⁵ to 1.6 × 10⁹ cells/mL. E. coli-imprinted OSX polymers with polyethylene glycol (PEG) differentiated between the targeted bacterium E. coli, and non-targeted bacteria S. typhimurium and native E. coli-GFP, achieving selectivity ratios up to 4.5 times higher than polydimethylsiloxane (PDMS) and OSX polymers without PEG.