Single polymeric microfiber waveguide platform for sensitive detection and discrimination of DNA methylation

Area of Science:

• Biotechnology
• Molecular Biology
• Analytical Chemistry

Background:

• Sensitive detection of DNA and DNA methylation is vital for biological research and disease diagnosis.
• Existing methods often involve complex procedures like PCR amplification.
• There is a need for rapid, sensitive, and user-friendly detection platforms for DNA methylation analysis.

Purpose of the Study:

• To develop a rapid and sensitive detection platform for p16 DNA and its methylation.
• To utilize a polymeric fluorescent microfiber waveguide with sandwich-structured hybridization for detection.
• To enable detection in complex biological samples like serum.

Main Methods:

• Conjugating oligonucleotides on microfiber surfaces and gold nanoparticles to capture target p16 DNA.
• Utilizing the fluorescence quenching effect upon target binding.
• Employing HpaII restriction enzyme digestion to differentiate methylated and unmethylated DNA.
• Monitoring the recovery of fluorescence signal after digestion to quantify methylation levels.

Main Results:

• The platform demonstrated selective and sensitive detection of p16 DNA with a limit of detection as low as 2 pM.
• The method achieved excellent analytical performance for detecting DNA methylation differences as low as 5%.
• The platform successfully detected p16 and its methylation in both buffer and serum samples.

Conclusions:

• The developed polymeric microfiber waveguide platform provides a sensitive and selective method for DNA and DNA methylation detection.
• This approach bypasses the need for traditional PCR amplification and simplifies operative processes.
• The technique holds significant potential for various DNA methylation analyses and early clinical disease diagnosis.