Development of a PCR-free electrochemical point of care test for clinical detection of methicillin resistant Staphylococcus aureus (MRSA)

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

Research output: Contribution to journalArticle

18 Citations (Scopus)

Abstract

An MRSA assay requiring neither labeling nor amplification of target DNA has been developed. Sequence specific binding of fragments of bacterial genomic DNA is detected at femtomolar concentrations using electrochemical impedance spectroscopy (EIS). This has been achieved using systematic optimisation of probe chemistry (PNA self-assembled monolayer film on gold electrode), electrode film structure (the size and nature of the chemical spacer) and DNA fragmentation, as these are found to play an important role in assay performance. These sensitivity improvements allow the elimination of the PCR step and DNA labeling and facilitate the development of a simple and rapid point of care test for MRSA. Assay performance is then evaluated and specific direct detection of the MRSA diagnostic mecA gene from genomic DNA, extracted directly from bacteria without further treatment is demonstrated for bacteria spiked into saline (106 cells per mL) on gold macrodisc electrodes and into human wound fluid (104 cells per mL) on screen printed gold electrodes. The latter detection level is particularly relevant to clinical requirements and point of care testing where the general threshold for considering a wound to be infected is 105 cells per mL. By eliminating the PCR step typically employed in nucleic acid assays, using screen printed electrodes and achieving sequence specific discrimination under ambient conditions, the test is extremely simple to design and engineer. In combination with a time to result of a few minutes this means the assay is well placed for use in point of care testing. 

LanguageEnglish
Pages6997-7005
Number of pages9
JournalAnalyst
Volume138
Issue number22
DOIs
Publication statusPublished - 21 Nov 2013
Externally publishedYes

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Point-of-Care Systems
Methicillin
Methicillin-Resistant Staphylococcus aureus
Assays
Electrodes
electrode
DNA
assay
Polymerase Chain Reaction
Gold
gold
Labeling
Bacteria
genomics
Dielectric Spectroscopy
Bacterial DNA
Intergenic DNA
bacterium
Nucleic acids
Wounds and Injuries

Cite this

Corrigan, D. K. ; Schulze, H. ; Henihan, G. ; Hardie, A. ; Ciani, I. ; Giraud, G. ; Terry, J. G. ; Walton, A. J. ; Pethig, R. ; Ghazal, P. ; Crain, J. ; Campbell, C. J. ; Templeton, K. E. ; Mount, A. R. ; Bachmann, T. T. / Development of a PCR-free electrochemical point of care test for clinical detection of methicillin resistant Staphylococcus aureus (MRSA). In: Analyst. 2013 ; Vol. 138, No. 22. pp. 6997-7005.
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Corrigan, DK, Schulze, H, Henihan, G, Hardie, A, Ciani, I, Giraud, G, Terry, JG, Walton, AJ, Pethig, R, Ghazal, P, Crain, J, Campbell, CJ, Templeton, KE, Mount, AR & Bachmann, TT 2013, 'Development of a PCR-free electrochemical point of care test for clinical detection of methicillin resistant Staphylococcus aureus (MRSA)' Analyst, vol. 138, no. 22, pp. 6997-7005. https://doi.org/10.1039/c3an01319g

Development of a PCR-free electrochemical point of care test for clinical detection of methicillin resistant Staphylococcus aureus (MRSA). / Corrigan, D. K.; Schulze, H.; Henihan, G.; Hardie, A.; Ciani, I.; Giraud, G.; Terry, J. G.; Walton, A. J.; Pethig, R.; Ghazal, P.; Crain, J.; Campbell, C. J.; Templeton, K. E.; Mount, A. R.; Bachmann, T. T.

In: Analyst, Vol. 138, No. 22, 21.11.2013, p. 6997-7005.

Research output: Contribution to journalArticle

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AU - Hardie, A.

AU - Ciani, I.

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