title: An Electrochemical DNA Sensing System Using Modified Nanoparticle Probes for Detecting Methicillin-Resistant Staphylococcus aureus.
abstract: We have developed a novel, highly sensitive, biosensing system for detecting methicillin-resistant Staphylococcus aureus (MRSA). The system employs gold nanoparticles (AuNPs), magnetic nanoparticles (mNPs), and an electrochemical detection method. We have designed and synthesized ferrocene- and single-stranded DNA-conjugated nanoparticles that hybridize to MRSA DNA. Hybridized complexes are easily separated by taking advantage of mNPs. A current response could be obtained through the oxidation of ferrocene on the AuNP surface when a constant potential of +250 mV vs. Ag/AgCl is applied. The enzymatic reaction of L-proline dehydrogenase provides high signal amplification. This sensing system, using a nanoparticle-modified probe, has the ability to detect 10 pM of genomic DNA from MRSA without amplification by the polymerase chain reaction. Current responses are linearly related to the amount of genomic DNA in the range of 10-166 pM. Selectivity is confirmed by demonstrating that this sensing system could distinguish MRSA from Staphylococcus aureus (SA) DNA.
authors.affiliation: Tenure-Track Program for Innovation Research, University of Fukui, Fukui, Japan.
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, Bunkyo 3-9-1, Fukui, 910-8507, 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.
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, Bunkyo 3-9-1, Fukui, 910-8507, Japan. suyeb10@u-fukui.ac.jp.|Department of Applied Chemistry and Biotechnology, Graduate School of Engineering, University of Fukui, Fukui, Japan. suyeb10@u-fukui.ac.jp.
authors.forename: Shin-Ichiro
authors.initials: SI
authors.lastname: Suye
mesh_terms: D015374:Biosensing Techniques; D002845:Chromatography; D004269:DNA, Bacterial / Q000032:analysis*; D055664:Electrochemical Techniques; D006046:Gold / Q000737:chemistry; D053768:Metal Nanoparticles; D055624:Methicillin-Resistant Staphylococcus aureus / Q000235:genetics*; D015335:Molecular Probes; D053758:Nanoparticles / Q000737:chemistry; D009693:Nucleic Acid Hybridization / Q000379:methods; D011394:Proline Oxidase / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins
chemical_list: D004269:DNA, Bacterial; D015335:Molecular Probes; D011994:Recombinant Proteins; D006046:Gold; D011394:Proline Oxidase
keywords: Chronoamperometry; DNA biosensor; Electrochemical detection; Magnetic separation; Nanoparticle