Rapid and sensitive detection of bacterial pathogens is critical for assessing public health, food and
environmental safety. We report the use of modularly designed and site-specifically oriented synthetic
antimicrobial peptides (sAMPs) as novel recognition agents enabling detection and quantification of
bacterial pathogens. The oriented assembly of the synthetic peptides on electrode surfaces through an
engineered cysteine residue coupled with impedimetric detection facilitated rapid and sensitive detection
of bacterial pathogens with a detection limit of 102 CFU/mL for four bacterial strains including Escherichia
coli (E. coli), Pseudomonas aeruginosa (P. aeruginosa), Staphylococcus aureus (S. aureus) and
Staphylococcus epidermidis (S. epidermidis). The approach enabled differentiation between live and dead
bacteria. The fabrication of the sAMPs functionalized surface and the importance of the sAMPs orientation
for providing optimum recognition and detection ability against pathogens are discussed. The
proposed methodology provides a universal platform for the detection of bacterial pathogens based on
engineered peptides, as alternative to the most commonly used immunological and gene based assays.
The method can also be used to fabricate antimicrobial coatings and surfaces for inactivation and
screening of viable bacteria.