Impedimetric detection of single-stranded PCR products derived from methicillin resistant Staphylococcus aureus (MRSA) isolates

D.K. Corrigan, H. Schulze, G. Henihan, I. Ciani, G. Giraud, J.G. Terry, A.J. Walton, R. Pethig, P. Ghazal, J. Crain, C.J. Campbell, A.R. Mount, T.T. Bachmann

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Using electrochemical impedance spectroscopy (EIS) the sensitive and specific detection of the antibiotic resistance gene mecA has been demonstrated. The gene sequence was obtained from clinical Staphylococcus aureus isolates. Initially a mecA specific probe was selected from hybridisation tests with a 3′ and 5′ version of a previously published probe sequence. When immobilised on a gold electrode in PNA form it was possible to detect hybridisation of mecA PCR product electrochemically at concentrations as low as 10 nM. By incorporating an undecane-thiol and 1.8 nm glycol spacer into the PNA probe it was possible to extend the limit of detection for mecA to 10 pM. Most published studies on EIS and nucleic acid detection report the use of short artificial DNA sequences or novel signal amplification schemes which improve sensitivity whereas this study reports the successful detection of long DNA fragments produced by PCR following extraction from clinical isolates. Finally, using screen printed electrodes the paper demonstrates hybridisation monitoring of mecA in an “on-line” assay format under ambient conditions which paves the way for rapid mecA detection in point of care scenarios.
LanguageEnglish
Pages178-184
Number of pages7
JournalBiosensors and Bioelectronics
Volume34
Issue number1
DOIs
Publication statusPublished - 15 Apr 2012

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Dielectric Spectroscopy
Methicillin
Methicillin-Resistant Staphylococcus aureus
Electrodes
Point-of-Care Systems
Electrochemical impedance spectroscopy
Polymerase Chain Reaction
Glycols
Microbial Drug Resistance
Genes
Sulfhydryl Compounds
Gold
Nucleic Acids
Staphylococcus aureus
Limit of Detection
DNA sequences
Nucleic acids
Antibiotics
Amplification
Assays

Keywords

  • electrochemical impedance spectroscopy
  • MRSA
  • mecA PCR product

Cite this

Corrigan, D.K. ; Schulze, H. ; Henihan, G. ; Ciani, I. ; Giraud, G. ; Terry, J.G. ; Walton, A.J. ; Pethig, R. ; Ghazal, P. ; Crain, J. ; Campbell, C.J. ; Mount, A.R. ; Bachmann, T.T. / Impedimetric detection of single-stranded PCR products derived from methicillin resistant Staphylococcus aureus (MRSA) isolates. In: Biosensors and Bioelectronics. 2012 ; Vol. 34, No. 1. pp. 178-184.
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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) isolates' Biosensors and Bioelectronics, vol. 34, no. 1, pp. 178-184. https://doi.org/10.1016/j.bios.2012.01.040

Impedimetric detection of single-stranded PCR products derived from methicillin resistant Staphylococcus aureus (MRSA) isolates. / Corrigan, D.K.; Schulze, H.; Henihan, G.; Ciani, I.; Giraud, G.; Terry, J.G.; Walton, A.J.; Pethig, R.; Ghazal, P.; Crain, J.; Campbell, C.J.; Mount, A.R.; Bachmann, T.T.

In: Biosensors and Bioelectronics, Vol. 34, No. 1, 15.04.2012, p. 178-184.

Research output: Contribution to journalArticle

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T1 - Impedimetric detection of single-stranded PCR products derived from methicillin resistant Staphylococcus aureus (MRSA) isolates

AU - Corrigan, D.K.

AU - Schulze, H.

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AU - Giraud, G.

AU - Terry, J.G.

AU - Walton, A.J.

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AU - Ghazal, P.

AU - Crain, J.

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