Iso-E-Codelock: A Rebuilding-free Electrochemical Chip with a Customizable Decoding Probe for Real-Time and Portable...

Area of Science:

• Biotechnology
• Analytical Chemistry
• Molecular Diagnostics

Background:

• Portable pathogen diagnostics require improved quantitation, digitization, and integration.
• Existing methods for isothermal nucleic acid amplification detection can be complex and lack real-time capabilities.

Purpose of the Study:

• To develop a ready-to-use electrochemical sensing strategy for real-time detection of isothermal nucleic acid amplification.
• To create a portable, sensitive, and selective diagnostic tool for pathogen identification.

Main Methods:

• Developed an electrochemical sensing strategy (Iso-E-Codelock) utilizing branched DNA as a codelock.
• Transduced isothermal amplicons into increased electrochemical probe current.
• Employed a self-designed portable instrument (BioAlex PHE-T) with a multichannel microchip for detection.
• Demonstrated the system with targets including SARS-CoV-2, African swine fever, FluA, and FluB genes.

Main Results:

• Achieved high sensitivity, high selectivity, signal-on response, and "zero" background.
• Enabled one-pot operation and real-time amplification curve output comparable to commercial PCR machines.
• The microchip is rebuilding-free and disposable.
• Customizable branch codelock probes demonstrated flexibility for different targets.

Conclusions:

• The Iso-E-Codelock strategy provides a high-performance, flexible, and portable solution for real-time pathogen detection.
• This approach simplifies quantitation, digitization, and integration in diagnostic applications.
• The system shows significant potential for widespread use in infectious disease diagnostics.