Rapid and Robust Semi-Qualitative Detection of DNA Using Nile Blue, A Visible Region Cationic Dye
Tech ID: 21-040
Inventors: Heather Whitehead, Marya Lieberman
Date Added: April 27, 2021
A field-friendly DNA quantification method
Rapid point-of-care (POC) tests are often desired for identifying viral, bacterial, and fungal pathogens in remote locations, where laboratory facilities are unavailable, and resources limited. Nucleic acid detection can facilitate quick and accurate diagnoses but requires equipment necessary to amplify the nucleic acids in patient samples to a detectable range. Further, current POC testing typically relies on UV-visualization techniques, which also require expensive, complex instrumentation. Finally, the reagents necessary for nucleic acid amplification are incompatible/interfere with UV-Vis DNA quantification, which requires an intermediary step of electrophoresis to purify the nucleic acid samples. Thus, there exists a need for an instrument-free detection method that can quickly determine and visualize the concentration of DNA present without additional tools, complex devices, or power.
Researchers at the University of Notre Dame have developed a rapid, robust, instrument-free colorimetric detection method for nucleic acids utilizing a cationic benzo phenoxazine dye that binds well with nucleic acids and is often used for DNA quantification. Changes in color intensities that occur when the dye binds to the DNA are correlated to the concentration of double-stranded DNA. This detection method allows quick and safe determination of the concentration of DNA by using a smartphone camera and free image analysis software and hazardous solvent-free methods. It only requires 20 uL of solution and tolerates various agents like phenol, chloroform, and ethanol. Additionally, this field-friendly candidate has minimal preparation and storage requirements. While this test can be used for diagnostic purposes in a field setting, it can also be useful for routine DNA amplification steps in a lab setting.
- Low power consumption
- Cheaper manufacturing
- Simple preparation and storage
- Safer- The dye remains stable for long periods of time
- Quick measurements
- Low resource alternative
- DNA amplification in lab
Technology Readiness Level
TRL 3 – Experimental Proof of Concept