Affinity capture of uropathogenic Escherichia coli using pigeon ovalbumin-bound Fe(3)O(4)@Al(2)O(3) magnetic nanoparticles

Abstract

Escherichia coli and Staphylococcus saprophyticus are the most common causes of urinary tract infections, with 80% of these infections caused by uropathogenic E. coli. Because the P fimbriae of E. coli have specificity toward Gal(alpha 1-4)Gal beta units, pigeon ovalbumin (POA), whose structure contains terminal Gal(alpha 1-4)Gal beta moieties, was used as a probe for interaction with P fimbriated E. coli. The functional affinity probes for these bacteria by immobilizing POA-a phosphoprotein-onto the surface of magnetic iron oxide nanoparticles (NPs) coated with alumina (Fe(3)O(4)@Al(2)O(3)), using the phosphate units of POA as linking groups for the formation of phosphate-alumina complexes. The immobilization process occurred within 30 s when performing the reaction under microwave heating. The magnetic POA-Fe(3)O(4)@Al(2)O(3) NPs generated using this facile approach exhibited specificity toward P fimbriated E. coli. The bacteria targeted by the affinity probes were characterized by matrix-assisted laser desorption/ionization mass spectrometry. The detection limit toward uropathogenic bacteria when using this approach was similar to 9.60 x 10(4) cfu/mL (0.5 mL).