ABSTRACT
Correction for 'Impedimetric measurement of DNA-DNA hybridisation using microelectrodes with different radii for detection of methicillin resistant Staphylococcus aureus (MRSA)' by Poh Quan Li et al., Analyst, 2017, 142, 1946-1952.
ABSTRACT
Due to their electroanalytical advantages, microelectrodes are a very attractive technology for sensing and monitoring applications. One highly important application is measurement of DNA hybridisation to detect a wide range of clinically important phenomena, including single nucleotide polymorphisms (SNPs), mutations and drug resistance genes. The use of electrochemical impedance spectroscopy (EIS) for measurement of DNA hybridisation is well established for large electrodes but as yet remains relatively unexplored for microelectrodes due to difficulties associated with electrode functionalisation and impedimetric response interpretation. To shed light on this, microelectrodes were initially fabricated using photolithography and characterised electrochemically to ensure their responses matched established theory. Electrodes with different radii (50, 25, 15 and 5 µm) were then functionalised with a mixed film of 6-mercapto-1-hexanol and a thiolated single stranded DNA capture probe for a specific gene from the antibiotic resistant bacterium MRSA. The complementary oligonucleotide target from the mecA MRSA gene was hybridised with the surface tethered ssDNA probe. The EIS response was evaluated as a function of electrode radius and it was found that charge-transfer (RCT) was more significantly affected by hybridisation of the mecA gene than the non-linear resistance (RNL) which is associated with the steady state current. The discrimination of mecA hybridisation improved as electrode radius reduced with the RCT component of the response becoming increasingly dominant for smaller radii. It was possible to utilise these findings to produce a real time measurement of oligonucleotide binding where changes in RCT were evident one minute after nanomolar target addition. These data provide a systematic account of the effect of microelectrode radius on the measurement of hybridisation, providing insight into critical aspects of sensor design and implementation for the measurement of clinically important DNA sequences. The findings open up the possibility of developing rapid, sensitive DNA based measurements using microelectrodes.
Subject(s)
Methicillin-Resistant Staphylococcus aureus/isolation & purification , Microelectrodes , Nucleic Acid Hybridization , DNA Probes , DNA, Bacterial , Genes, BacterialABSTRACT
The performance of two electrode architectures with broadly similar overall active electrode areas are examined. The first is an electrode comprising a single contiguous area (a disc) and the second is an electrode in which the cumulative electrode area is dispersed over a wide area as a 50 nm thickness platinum nanoband. A direct comparison of the electrochemical performance of these two electrodes has been made. The relatively simple nanoband electrode architecture is shown to have benefits, including two orders of magnitude greater mass transport limited currents, the ability to measure faster electrode kinetics (by a similar factor), a three orders of magnitude lowering of the Limit of Detection and a significantly reduced susceptibility to hydrodynamic perturbations. The consequences and implications of these performance characteristics on the uses of such a nanoband electrode have been considered.
ABSTRACT
Micron resolution photolithography has been employed to make microsquare nanoband edge electrode (MNEE) arrays with reproducible and systematic control of the crucial dimensional parameters, including array element size and spacing and nanoelectrode thickness. The response of these arrays, which can be reproducibly fabricated on a commercial scale, is first established. The resulting characteristics (including high signal and signal-to-noise, low limit of detection, insensitivity to external convection and fast, steady-state, reproducible and quantitative response) make such nanoband electrode arrays of real interest as enhanced electroanalytical devices. In particular, the nanoelectrode response is presented and analysed as a function of nanometre scale electrode dimension, to assess the impact and relative contributions of previously postulated nanodimensional effects on the resulting response. This work suggests a significant contribution of migration at the band edges to mass transfer, which affects the resulting electroanalytical response even at ionic strengths as large as 0.7 mol dm(-3) and for electrodes as wide as 50 nm. For 5 nm nanobands, additional nanoeffects, which are thought to arise from the fact that the size of the redox species is comparable to the band width, are also observed to attenuate the observed current. The fundamental insight this gives into electrode performance is discussed along with the consequent impact on using such electrodes of nanometre dimension.