Label- and amplification-free electrochemical detection of bacterial ribosomal RNA
Henihan, Grace and Schulze, Holger and Corrigan, Damion K. and Giraud, Gerard and Terry, Jonathan G. and Hardie, Alison and Campbell, Colin J. and Walton, Anthony J. and Crain, Jason and Pethig, Ronald and Templeton, Kate E. and Mount, Andrew R. and Bachmann, Till T. (2016) Label- and amplification-free electrochemical detection of bacterial ribosomal RNA. Biosensors and Bioelectronics, 81. pp. 487-494. ISSN 0956-5663 (https://doi.org/10.1016/j.bios.2016.03.037)
Preview |
Text.
Filename: Henihan_etal_BB_2016_label_and_amplification_free_electrochemical_detection_of_bacterial_ribosomal_RNA.pdf
Accepted Author Manuscript License: Download (871kB)| Preview |
Abstract
Current approaches to molecular diagnostics rely heavily on PCR amplification and optical detection methods which have restrictions when applied to point of care (POC) applications. Herein we describe the development of a label-free and amplification-free method of pathogen detection applied to Escherichia coli which overcomes the bottleneck of complex sample preparation and has the potential to be implemented as a rapid, cost effective test suitable for point of care use. Ribosomal RNA is naturally amplified in bacterial cells, which makes it a promising target for sensitive detection without the necessity for prior in vitro amplification. Using fluorescent microarray methods with rRNA targets from a range of pathogens, an optimal probe was selected from a pool of probe candidates identified in silico. The specificity of probes was investigated on DNA microarray using fluorescently labeled 16S rRNA target. The probe yielding highest specificity performance was evaluated in terms of sensitivity and a LOD of 20 pM was achieved on fluorescent glass microarray. This probe was transferred to an EIS end point format and specificity which correlated to microarray data was demonstrated. Excellent sensitivity was facilitated by the use of uncharged PNA probes and large 16S rRNA target and investigations resulted in an LOD of 50 pM. An alternative kinetic EIS assay format was demonstrated with which rRNA could be detected in a species specific manner within 10-40 min at room temperature without wash steps.
ORCID iDs
Henihan, Grace, Schulze, Holger, Corrigan, Damion K. ORCID: https://orcid.org/0000-0002-4647-7483, Giraud, Gerard, Terry, Jonathan G., Hardie, Alison, Campbell, Colin J., Walton, Anthony J., Crain, Jason, Pethig, Ronald, Templeton, Kate E., Mount, Andrew R. and Bachmann, Till T.;-
-
Item type: Article ID code: 56727 Dates: DateEvent15 July 2016Published18 March 2016Published Online17 March 2016AcceptedSubjects: Technology > Engineering (General). Civil engineering (General) > Bioengineering Department: UNSPECIFIED Depositing user: Pure Administrator Date deposited: 23 Jun 2016 08:59 Last modified: 22 Sep 2024 00:50 Related URLs: URI: https://strathprints.strath.ac.uk/id/eprint/56727