A sensitive ligase-based ATP electrochemical assay using molecular beacon-like DNA

Area of Science:

• Electrochemistry
• Molecular Biology
• Biosensing

Background:

• Adenosine-5'-triphosphate (ATP) is a vital molecule in cellular energy transfer.
• Accurate ATP detection is essential for various biological and environmental applications.
• Existing detection methods may lack sensitivity, selectivity, or require complex procedures.

Purpose of the Study:

• To develop a sensitive and selective signal-on electrochemical sensing method for ATP detection.
• To utilize a molecular beacon (MB)-like DNA strategy combined with ligase activity.
• To establish a robust platform for ATP quantification in complex samples.

Main Methods:

• Self-assembly of biotin-tagged MB-like DNA onto a gold electrode via gold-thiol chemistry.
• ATP-dependent ligation of DNA fragments by T4 DNA ligase, triggering a conformational change.
• Electrochemical signal generation measured by differential pulse voltammetry.

Main Results:

• Achieved sensitive ATP detection down to 0.05 nM with a linear response from 0.1 to 1000 nM.
• Demonstrated high selectivity, discriminating ATP from its analogues.
• Successfully applied the method for ATP determination in Escherichia coli O157:H7 extracts from water samples.

Conclusions:

• The developed ligase-based electrochemical sensor provides a sensitive and selective platform for ATP detection.
• The signal-on approach offers a reliable method for quantifying ATP in real-world samples.
• This method holds promise for applications in environmental monitoring and diagnostics.