RGB colour coding of Y-shaped DNA for simultaneous tri-analyte solid phase hybridization detection

Area of Science:

• Molecular Biology
• Biotechnology
• Analytical Chemistry

Background:

• Multiplex DNA detection is crucial for identifying multiple pathogens simultaneously.
• Existing methods can be complex and time-consuming.

Purpose of the Study:

• To develop a novel, rapid, and color-coded method for tri-analyte DNA detection.
• To establish a proof-of-concept assay for detecting specific pathogenic bacterial DNA sequences.

Main Methods:

• Utilized a Y-shaped capture probe design for simultaneous binding of three DNA targets.
• Employed fluorescently labeled reporter probes to generate an RGB color-coded output.
• Developed a method to correlate RGB values with target DNA concentration ratios.

Main Results:

• Successfully demonstrated semi-quantitative detection of DNA sequences from *Escherichia coli* O157:H7, *Vibrio cholera*, and *Salmonella enteric*.
• Achieved detection across a concentration range of 20-167 nM.
• The assay provided data interpretation within approximately 4 hours from hybridization.

Conclusions:

• The Y-shaped probe assay offers a novel approach for rapid, multiplexed DNA detection.
• The RGB color-coding system provides a visual and quantitative relationship to target DNA ratios.
• This method shows potential for quick identification of multiple bacterial pathogens.