ArticleApplied biochemistry and biotechnology2026
Enzyme-Free DNA Logic Circuit with Single-Color Readout for Dual Biomarker Detection.
Article in Applied biochemistry and biotechnology, 2026. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Not yet cited in PubMed.
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Abstract
This study introduces an innovative enzyme-free DNA circuit system for the simultaneous detection of two distinct nucleic acid biomarkers using a single colorimetric output with four discrete intensity levels. The system integrates toehold-mediated strand displacement (TMSD) and catalytic hairpin assembly (CHA) to generate diagnostic signals based on G-quadruplex structures. Two logic gates, ABC and DE, are designed to respond differentially to inputs: the ABC gate disassembles two G-quadruplexes upon recognition of Input 1, while the DE gate forms a G-quadruplex in response to Input 2. This configuration yields four unique output states corresponding to distinct input combinations. The circuit achieves high sensitivity, with detection limits of 5 pM for Input 1 and 1 pM for Input 2, and displays strong specificity against non-target DNA sequences. Validation in complex biological matrices confirmed the robustness of the system: it maintained 85 ± 3% signal in 50% human serum, indicating high compatibility with clinical sample environments. By employing a single output signal format, the approach simplifies data interpretation, reduces processing time, and enhances cost-effectiveness compared to conventional multi-signal platforms. As an automated, enzyme-free, and one-step detection system, this platform offers a powerful tool for accurate and efficient multiplex biomarker analysis, with promising applications in early disease diagnosis and point-of-care testing.
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