Small Molecule Detection with Ligation-Dependent Light-Up Aptamer Transcriptional Amplification

Area of Science:

• Biochemistry
• Molecular Biology
• Biotechnology

Background:

• Adenosine triphosphate (ATP) and nicotinamide adenine dinucleotide (NAD+) are crucial biomolecules involved in numerous physiological processes.
• These molecules are recognized as potential biomarkers for diagnosing various diseases.

Purpose of the Study:

• To develop a sensitive and selective assay for detecting ATP and NAD+.
• To achieve significant signal amplification for improved biomarker detection.

Main Methods:

• A ligation-dependent light-up aptamer transcriptional amplification assay was developed.
• The assay utilizes a specific DNA ligase to ligate a nicked DNA template in the presence of the target molecule.
• The ligation product undergoes in vitro transcription to amplify fluorescence signals mediated by a light-up aptamer.

Main Results:

• The assay achieved low detection limits: 5.9 pM for ATP and 142 pM for NAD+.
• High selectivity was demonstrated, distinguishing targets from similar molecules.
• The method showed excellent recovery rates when applied to human serum samples.

Conclusions:

• The developed assay provides a sensitive and selective platform for detecting ATP and NAD+.
• This method holds potential for biomarker-based disease diagnosis.
• The assay's effectiveness in biological samples highlights its practical applicability.