Electrochemical detection of synthetic DNA and native 16S rRNA fragments on a microarray using a biotinylated intercalator as coupling site for an enzyme label

Zimdars, Andreas; Gębala, Magdalena; Hartwich, Gerhard; Neugebauer, Sebastian; Schuhmann, Wolfgang

doi:10.1016/j.talanta.2015.04.041

Cited by 13 publications

(16 citation statements)

References 29 publications

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“…Liao et al and Elsholz et al reported LODs of 4 x 10 4 CFU/mL and 10 5 CFU/mL (0.5 ng/µL total RNA) which are slightly higher than the LOD obtained in this study (Elsholz et al 2006;Liao et al 2006). Zimdars et al recently reported a very low LOD of 30 fM 16S rRNA using a biotinylated intercalator as a coupling site for a streptavidin/alkaline phosphatase enzyme label (Zimdars et al 2015). But these enzyme label based amperometric biosensors have several disadvantages regarding POCT development.…”

Section: Electrochemical Impedance Spectroscopy Kinetic Measurementsmentioning

confidence: 99%

“…The research group of Shana O. Kelley used signal amplification based on Ru 3+ adsorption towards hybridized target nucleic acids and Fe 3+ amplification at the surface of nanostructured microelectrodes for the direct detection of E. coli mRNA by differential pulse voltammetry and combined this with electrochemical lysis of bacterial cells (Besant et al 2013;Sage et al 2014). Several groups applied enzymatic signal amplification using Streptavidin-conjugated horseradish peroxidase or alkaline phosphatase bound to the hybridized 16S rRNA via biotinylated detection probes or biotinylated intercalator for the amperometric detection of bacterial RNA (Elsholz et al 2006;Gau et al 2001;Liao et al 2006;Liao et al 2007;Sin et al 2013;Williams et al 2003;Zimdars et al 2015). All these enzymatic signal amplification methods require multiple incubation and washing steps which makes them less amenable to implementation in a fully integrated sample-to-answer system.…”

Section: Introductionmentioning

confidence: 99%

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Label- and amplification-free electrochemical detection of bacterial ribosomal RNA

Henihan

Schulze

Corrigan

et al. 2016

Biosensors and Bioelectronics

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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 10-40min at room temperature without wash steps.

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Section: Electrochemical Impedance Spectroscopy Kinetic Measurementsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Label- and amplification-free electrochemical detection of bacterial ribosomal RNA

Henihan

Schulze

Corrigan

et al. 2016

Biosensors and Bioelectronics

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“…Zimdars et al utilized a detection scheme with a DNA intercalator, which selectively bound to the DNA probe if it had hybridized with a target sequence. Assays were performed on 32 individually addressable microelectrodes and the lowest measured concentration of RNA was 30 fM and 60 nM of synthetic DNA (Zimdars et al, 2015). .…”

Section: Microelectrodesmentioning

confidence: 99%

A review of microfabricated electrochemical biosensors for DNA detection

Blair

Corrigan

2019

Biosensors and Bioelectronics

130

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This review article presents an overview of recent work on electrochemical biosensors developed using microfabrication processes, particularly sensors used to achieve sensitive and specific detection of DNA sequences. Such devices are important as they lend themselves to miniaturisation, reproducible mass-manufacture, and integration with other previously existing technologies and production methods. The review describes the current state of these biosensors, novel methods used to produce them or enhance their sensing properties, and pathways to deployment of a complete point-ofcare biosensing system in a clinical setting.

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“…Biosensors based on electrochemical read‐outs offer several advantages for their implementation as POC devices, including their inherent high sensitivity, low cost, and potential for miniaturization . Therefore, considerable effort has been put into developing electrochemical diagnostic platforms capable of rapid biomarker identification and pathogen detection .…”

Section: Introductionmentioning

confidence: 99%

Electric Field Modulation of Silicon upon Tethering of Highly Charged Nucleic Acids. Capacitive Studies on DNA‐modified Silicon (111)

et al. 2016

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Electrochemical detection of synthetic DNA and native 16S rRNA fragments on a microarray using a biotinylated intercalator as coupling site for an enzyme label

Cited by 13 publications

References 29 publications

Label- and amplification-free electrochemical detection of bacterial ribosomal RNA

Label- and amplification-free electrochemical detection of bacterial ribosomal RNA

A review of microfabricated electrochemical biosensors for DNA detection

Electric Field Modulation of Silicon upon Tethering of Highly Charged Nucleic Acids. Capacitive Studies on DNA‐modified Silicon (111)

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