title: A smart DNA sensing system for detecting methicillin-resistant Staphylococcus aureus using modified nanoparticle probes.
abstract: Methicillin-resistant Staphylococcus aureus (MRSA) is one of the most common causes of hospital-acquired infections. To prevent epidemics, a quick and simple detection method is required. In this study, we developed a novel electrochemical DNA detection method that does not rely upon polymerase chain reaction (PCR) and may be used in point-of-care facilities. The electrochemical DNA sensing system presented here is based on the chronoamperometric detection of ferrocene-labeled probes that were conjugated to gold nanoparticles (AuNPs). This DNA sensor system employed magnetic nanoparticle (MNP)-modified probes allowing easy sample DNA recovery. AuNP nanoparticles with ferrocene-labeled probes enabled the generation of an electric signal, and MNP/DNA/AuNP conjugates were formed by hybridization. Following hybridization, the MNP/DNA/AuNP hybridization complex is magnetically separated, and electrochemical current responses could be obtained because of the AuNP-ferrocene complexes. To construct a highly sensitive system, dye-linked L-proline dehydrogenase (L-proDH) was employed to amplify current responses following a catalytic reaction with L-proline. Rapid catalytic reaction of L-proDH and substrate was able to amplify the oxidation of ferrocene. Target DNA from MRSA could be quantified over a range of 10-166pM, and this sensing system could also distinguish MRSA from S. aureus.
authors.affiliation: Department of Frontier Fiber Technology and Science, Graduate School of Engineering, University of Fukui, Fukui, Japan.
authors.forename: Kazuya
authors.initials: K
authors.lastname: Watanabe
authors.affiliation: Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, Fukui, Japan.
authors.forename: Noriaki
authors.initials: N
authors.lastname: Kuwata
authors.affiliation: Tenure-Track Program for Innovation Research, University of Fukui, Japan. Electronic address: hi-saka@u-fukui.ac.jp.
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Sakamoto
authors.affiliation: Department of Frontier Fiber Technology and Science, Graduate School of Engineering, University of Fukui, Fukui, Japan.
authors.forename: Yoshihisa
authors.initials: Y
authors.lastname: Amano
authors.affiliation: Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, Fukui, Japan. Electronic address: satomura@u-fukui.ac.jp.
authors.forename: Takenori
authors.initials: T
authors.lastname: Satomura
authors.affiliation: Department of Frontier Fiber Technology and Science, Graduate School of Engineering, University of Fukui, Fukui, Japan; Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, Fukui, Japan. Electronic address: suyeb10@u-fukui.ac.jp.
authors.forename: Shin-ichiro
authors.initials: S
authors.lastname: Suye
mesh_terms: D015374:Biosensing Techniques; D004269:DNA, Bacterial / Q000235:genetics / Q000302:isolation & purification*; D006046:Gold; D006801:Humans; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics / Q000302:isolation & purification*; D053758:Nanoparticles; D013203:Staphylococcal Infections / Q000175:diagnosis* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D006046:Gold
keywords: Chronoamperometry; DNA sensor; Magnetic separation; Methicillin-resistant Staphylococcus aureus; Nanoparticle