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.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
References
Corrigan DK, Schulze H, Henihan G, Ciani I, Giraud G, Terry JG, Walton AJ, Pethig R, Ghazal P, Crain J, Campbell CJ, Mount AR, Bachmann TT (2012) Impedimetric detection of single-stranded PCR products derived from methicillin resistant Staphylococcus aureus (MRSA) isolated. Biosens Bioelectron 34:178–184
Bessède E, Delcamp A, Sifré E, Buissonnière A, Mégraud F (2011) New methods for detection of campylobacters in stool samples in comparison to culture. J Clin Microbiol 49:941–944
Conza L, Casati S, Gaia V (2013) Detection limits of Legionella pneumophila in environmental samples after co-culture with Acanthamoeba polyphage. BMC Microbiol 13:49
Schabereiter-Gurtner C, Selitsch B, Rotter ML, Hirschl AM, Williger B (2007) Development of novel real-time PCR assays for detection and differentiation of eleven medically important Aspergillus and Candida species in clinical specimens. J Clin Microbiol 45:906–914
Lee DY, Shannon K, Beaudette LA (2006) Detection of bacterial pathogens in municipal wastewater using an oligonucleotide microarray and real-time quantitative PCR. J Microbiol Methods 65:453–467
Gu Y, Lai MT (2012) The potential application of a poly(3,4-ethylenedioxythiopene) modified platinum DNA biosensor in mutation analysis. Biosens Bioelectron 31:124–129
Ding Y, Wang Q, Gao F, Gao F (2013) Highly sensitive and selective DNA biosensor using dumbbell-shaped bis-groove binder of bi-acetylferrocene ethylenediamine complex as electrochemical indicator. Electrochim Acta 106:35–42
Li F, Han X, Liu S (2011) Development of an electrochemical DNA biosensor with a high sensitivity of fM by dendritic gold nanostructure modified electrode. Biosens Bioelectron 26:2619–2625
Watanabe K, Kuwata N, Sakamoto H, Amano Y, Satomura T, Suye S (2015) A smart DNA sensing system for detecting methicillin-resist Staphylococcus aureus using modified nanoparticle probes. Biosens Bioelectron 15:419–423
Satomura T, Hara Y, Suye S, Sakuraba H, Ohshima T (2012) Gene expression and characterization of a third type of dye-linked l-proline dehydrogenase from the aerobic hyperthermophilic archaeon, Aeropyrum pernix. Biosci Biotech Bioch 76:589–593
Acknowledgment
This is the author’s version of a work accepted for publication by Elsevier. Changes resulting from the publishing process, including peer review, editing, corrections, structural formatting, and other quality control mechanisms, may not be reflected in this document. Changes may have been made to this work since it was submitted for publication. The definitive version has been published in Biosensor and Bioelectronics, VOLUME 67, SPECIAL ISSUE BIOSENSORS 2014, May 15, DOI: 10.1016/j.bios.2014.08.075
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2017 Springer Science+Business Media LLC
About this protocol
Cite this protocol
Sakamoto, H., Amano, Y., Satomura, T., Suye, Si. (2017). An Electrochemical DNA Sensing System Using Modified Nanoparticle Probes for Detecting Methicillin-Resistant Staphylococcus aureus . In: Prickril, B., Rasooly, A. (eds) Biosensors and Biodetection. Methods in Molecular Biology, vol 1572. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6911-1_2
Download citation
DOI: https://doi.org/10.1007/978-1-4939-6911-1_2
Published:
Publisher Name: Humana Press, New York, NY
Print ISBN: 978-1-4939-6910-4
Online ISBN: 978-1-4939-6911-1
eBook Packages: Springer Protocols