A detection system using sensing motif-tethered oligodeoxynucleotides for multiplex biomolecular analysis

Area of Science:

• Biochemistry
• Molecular Biology
• Analytical Chemistry

Background:

• Oligodeoxynucleotides offer versatile platforms for molecular recognition.
• Developing sensitive detection systems for biomolecules like glutathione (GSH) and hydrogen peroxide (H2O2) is crucial in various biological and medical fields.
• Existing methods may lack specificity or require complex sample preparation.

Purpose of the Study:

• To develop and validate a novel DNA-based system for the simultaneous detection of multiple target biomolecules.
• To utilize sensing motif-tethered oligodeoxynucleotides as molecular probes.
• To enable sensitive quantification of target biomolecules through qPCR.

Main Methods:

• Oligodeoxynucleotides functionalized with sensing motifs were designed.
• The probes reacted with target biomolecules, glutathione (GSH) and hydrogen peroxide (H2O2), generating a primary amine motif.
• Biotin labeling was employed for selective capture of product DNAs.
• Quantitative Polymerase Chain Reaction (qPCR) was used for final quantification.

Main Results:

• The developed system successfully detected the presence of target biomolecules.
• The reaction with biomolecules yielded a detectable primary amine motif on the DNA probes.
• Selective capture and subsequent qPCR analysis allowed for sensitive quantification.

Conclusions:

• A novel, sensitive, and selective DNA-based detection system for multiple biomolecules was successfully developed.
• The system leverages the unique reactivity of sensing motif-tethered oligodeoxynucleotides.
• This approach offers a promising tool for biomolecular detection and quantification in various applications.