Antimicrobial Peptides as Probes for Electrochemical Detection of Methicillin-resistant Staphylococcus Aureus

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PROJECT SUMMARY/ABSTRACT The broad, long-term goal of Project Leader Rebecca Lai’s project is to develop a near real-time method for detecting methicillin-resistant Staphylococcus aureus (MRSA). In recent decades, MRSA strains have become the most common cause of health care-associated infections, and new multi-drug-resistant strains of this severe and potentially life-threatening disease have emerged. Although implementation of universal admission surveillance can reduce the spread of MRSA infections, it is not practiced in many health care systems because of the cost of testing. Thus, there is a critical need for more advanced detection methods that are sensitive, specific, and rapid yet cost-effective enough to be employed in hospitals and health care facilities in low resource settings. An area that deserves attention in MRSA sensor design is the selection of appropriate biorecognition elements. Among available biorecognition elements, antimicrobial peptides (AMPs), which are short peptide fragments existing in various forms of life, are one of the best alternatives to antibodies because of a combination of attractive properties: high stability, synthetic simplicity, easy accessibility, and high affinity toward their specific bacterial targets. Thus, the goal of this project is to evaluate and characterize MRSA-specific peptides suitable for use as biorecognition elements in the fabrication of E-PB MRSA sensors and to use potential-assisted “click” chemistry in the construction of a paper-based sensor array. Two aims will be achieved: 1) design and fabricate E-PB sensors using high efficacy AMPs for detection of MRSA and 2) develop a paper-based E-PB sensor array using potential-assisted “click” chemistry. To ensure that the sensor array will perform optimally, critical aspects of the bioconjugation reaction will be systematically evaluated, including the reaction time, applied potential, and concentrations of the reagents. This project will result in an E-PB sensor array capable of near real-time detection of MRSA bacterial cells, which will be an important step toward the development of an all-in-one microfluidic- based MRSA sensor device. Project outcomes will stimulate future research on AMPs for biosensing applications. The availability of such a versatile sensing platform could have a major impact on MRSA diagnosis, management, and surveillance. This project will also contribute to the wider goal of the Nebraska Center for Integrated Biomolecular Communication (CIBC), which includes the development of novel biomolecular sensing strategies for disease diagnosis. It will enhance CIBC’s research strengths in biosensing and bioanalysis and facilitate future development of sensors for other important diseases.