A Bacteriophage-Based Electrochemical Biosensor for Detection of Methicillin-Resistant Staphylococcus aureus

Abstract

Bacterial pathogens are important targets for detection and early diagnostics of infection in healthcare. Methicillin-resistant Staphylococcus aureus (MRSA) is a bacterium that can cause severe complications such as pneumonia and septicemia from surgical site infections. In this work, a carbon nanotube (CNT)-based impedimetric biosensor using SATA-8505, a bacteriophage, as a recognition element was developed for the detection of MRSA USA300, a common strain of MRSA found in surgical infections. The biosensor allows for fast and selective detection of viable MRSA cells by measuring impedance changes on the electrode surface as a result of bacterial capture by the phage-functionalized CNT electrode. The immobilization of the bacteriophage was achieved by inducing an electric-field and using a charge-directed orientation strategy, and activity of the immobilized phage was confirmed by an infectivity study using disk diffusion methods. The biosensing platform showed high selectivity and sensitivity toward MRSA USA300 with a limit of detection of 1.23 × 102 CFU/ml in aqueous solution and 1.29 × 102 CFU/ml in blood plasma. The biosensing platform could potentially be integrated into a lab-on-a-chip platform for point of care use and for other pathogens such as Pseudomonas aeruginosa, Klebsiella and Listeria monocytogenes.

Recent Developments in Phenotypic and Molecular Diagnostic Methods for Antimicrobial Resistance Detection in Staphylococcus aureus: A Narrative Review

“…The authors tested the system in 30 nasal samples and it correctly identified 11 MRSA and 19 non-MRSA isolates [ 123 ]. Patel and colleagues developed an electrochemical biosensor to detect one MRSA strain using the bacteriophage SATA-8505 [ 135 ]. Maldonado and co-authors developed a nanophotonic biosensor which uses a bimodal waveguide interferometer to differentiate MRSA from MSSA, based on PBP2a protein detection [ 136 ].…”

Electrochemical biosensors for rapid diagnosis of bacterial infections: Design, targets and applications in clinical setting

“…Their system was a carbon nanotube-based impedimetric biosensor with SATA-8505 bacteriophage as a recognition element. Their system showed high selectivity and sensitivity in blood samples [82].…”

A novel electrochemical sandwich-like immunosensor based on carboxyl Ti3C2Tx MXene and rhodamine b/gold/reduced graphene oxide for Listeria monocytogenes

“…As one of the most common foodborne pathogens, Listeria monocytogenes (LM), which can induce various serious diseases (e.g. meningitis, febrile gastroenteritis, abortion and septicemia) to human health, [1][2][3] and LM can survive easily in a series of foods including water, cheese, milk, meats and raw vegetables, etc. 4,5 In USA, a 0-tolerance policy was formed for the presence of LM in food; in Canada, only 100 CFU/g was allowed.…”

“…meningitis, febrile gastroenteritis, abortion and septicemia) to human health. 1–3 LM can survive easily in a series of foods including water, cheese, milk, meats and raw vegetables. 4,5 In the USA, a zero-tolerance policy was formed for the presence of LM in food; in Canada, only 100 CFU g −1 was allowed.…”