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Sensors 2018, 18(10), 3259; https://doi.org/10.3390/s18103259

Implementing Morpholino-Based Nucleic Acid Sensing on a Portable Surface Plasmon Resonance Instrument for Future Application in Environmental Monitoring

1
International Research Institute of Stavanger, 4068 Stavanger, Norway
2
Departments of Medicine-Division of Medical Genetics and Genome Sciences, University of Washington, Seattle, WA 98195-5852, USA
*
Author to whom correspondence should be addressed.
The International Research Institute of Stavanger will become part of (and renamed to): “NORCE Norwegian Research Center” from the first of October 2018.
Received: 3 September 2018 / Revised: 22 September 2018 / Accepted: 26 September 2018 / Published: 28 September 2018
(This article belongs to the Special Issue Surface Plasmon Resonance Sensing 2019)
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Abstract

A portable surface plasmon resonance (SPR) instrument was tested for the first time for the detection of oligonucleotide sequences derived from the 16S rRNA gene of Oleispira antarctica RB-8, a bioindicator species of marine oil contamination, using morpholino-functionalized sensor surfaces. We evaluated the stability and specificity of morpholino coated sensor surfaces and tested two signal amplification regimes: (1) sequential injection of sample followed by magnetic bead amplifier and (2) a single injection of magnetic bead captured oligo. We found that the sensor surfaces could be regenerated for at least 85 consecutive sample injections without significant loss of signal intensity. Regarding specificity, the assay clearly differentiated analytes with only one or two mismatches. Signal intensities of mismatch oligos were lower than the exact match target at identical concentrations down to 200 nM, in standard phosphate buffered saline with 0.1 % Tween-20 added. Signal amplification was achieved with both strategies; however, significantly higher response was observed with the sequential approach (up to 16-fold), where first the binding of biotin-probe-labeled target oligo took place on the sensor surface, followed by the binding of the streptavidin magnetic beads onto the immobilized targets. Our experiments so far indicate that a simple coating procedure in combination with a relatively cost-efficient magnetic-bead-based signal amplification will provide robust SPR based nucleic acid sensing down to 0.5 nM of a 45-nucleotide long oligo target (7.2 ng/mL). View Full-Text
Keywords: surface plasmon resonance; 16S rRNA; bacterial detection; hydrocarbon biodegradation; magnetic bead based biosensing; portable biosensor surface plasmon resonance; 16S rRNA; bacterial detection; hydrocarbon biodegradation; magnetic bead based biosensing; portable biosensor
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This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited (CC BY 4.0).
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Bagi, A.; Soelberg, S.D.; Furlong, C.E.; Baussant, T. Implementing Morpholino-Based Nucleic Acid Sensing on a Portable Surface Plasmon Resonance Instrument for Future Application in Environmental Monitoring. Sensors 2018, 18, 3259.

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