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Lysostaphin-mediated cell fragmentation for sensitive label-free impedance immunoanalysis of Staphylococcus aureus

Park, Yoo Min; Han, Yong Duk; Chun, Hyeong Jin; Yoon, Hyun C.
Department of Otolaryngology, Ajou University School of Medicine
Staphylococcus aureus (S.aureus) is one of common and a lethal pathogen that may be found in the skin and mucous membranes of human. Infection of S.aureus through the wound of the skin such as acne or furuncle is primary cause of septicemia. Therefore, an accurate detection of the S.aureus is a significant issue in diagnosis of infectious disease. Based on this requirement, we developed an impedance biosensor for the S.aureus detection. Electrochemical impedance spectroscopy (EIS) is a sensitive and simple label-free analytical method used for measurement of the target-specific sensing surface by examining characteristics of the electrode. Compared to the previously reported electrochemical S.aureus biosensor which requires repetitive laborious cell-washing steps, our new system requires no cell-washing step except simple treatment of an enzyme (Lysostaphin) which specifically fragmentizes the cell wall of S.aureus. Lysostaphin rapidly and specifically cleaves the pentaglycine cross-bridges of the peptidoglycan on the S. aureus cell walls but has no effect on other bacteria. In our research, lysostaphin and its S. aureus-lytic activity were introduced to the EIS-based immunoassay for enhancement of the sensitivity of the assay.
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