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Bioluminescent Intercalating Dyes for Ratiometric Nucleic Acid Detection.
Yosta de Stigter1,2, Harmen J van der Veer1,2, Bas J H M Rosier1,2
1Laboratory of Chemical Biology, Department of Biomedical Engineering, Eindhoven University of Technology, 5600 MB Eindhoven, The Netherlands.
ACS Chemical Biology
|February 5, 2024
Summary
Researchers developed Luminescent Multivalent Intercalating Dye (LUMID) sensors for rapid, sensitive DNA detection. This novel bioluminescent tool enables point-of-need diagnostics without specialized equipment, improving disease monitoring capabilities.
Area of Science:
- Biotechnology
- Molecular Biology
- Diagnostics
Background:
- Current DNA detection methods often require specialized equipment and target-specific probes, hindering point-of-need applications.
- Developing rapid, sensitive, and broadly applicable DNA detection assays is crucial for disease diagnosis and monitoring.
Purpose of the Study:
- To develop a novel, off-the-shelf bioluminescent sensor for nonspecific double-stranded DNA detection.
- To enhance DNA detection sensitivity and specificity for point-of-need applications, such as viral DNA detection.
Main Methods:
- Conjugation of intercalating dyes to NanoLuc luciferase to create Luminescent Multivalent Intercalating Dye (LUMID) sensors.
- Incorporation of multiple, tandem-arranged dyes with positively charged linkers to improve DNA-binding affinity.
- Integration of LUMID with loop-mediated isothermal amplification (LAMP) for sequence-specific detection.
Main Results:
- LUMID sensors demonstrated nonspecific double-stranded DNA detection via a blue-to-green color change.
- Improved DNA-binding affinities by over 2 orders of magnitude, detecting nanomolar DNA concentrations with an 8-fold change in the green/blue ratio.
- Achieved attomolar sensitivity for viral DNA detection when combined with LAMP, enabling smartphone-based readout.
Conclusions:
- LUMID offers a simple, sensitive, and versatile tool for DNA detection.
- The developed sensor system is particularly suitable for point-of-need diagnostic applications.
- LUMID represents a significant advancement in accessible molecular diagnostics.
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