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Aptamer-Based Target Detection Facilitated by a 3-Stage G-Quadruplex Isothermal Exponential Amplification Reaction

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Label-Free, Visual Detection of Small Molecules Using Highly Target-Responsive Multimodule Split Aptamer Constructs.

Yingping Luo^1,2, Haixiang Yu¹, Obtin Alkhamis¹

¹Department of Chemistry and Biochemistry , Florida International University , 11200 SW Eighth Street , Miami , Florida 33199 , United States.

Analytical Chemistry

|May 4, 2019

Summary

This summary is machine-generated.

Researchers developed novel DNA sensors called CBSAzymes for rapid, visual detection of small molecules. These sensors enable naked-eye identification of substances like cocaine and synthetic cathinones in samples without instruments.

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Area of Science:

Biotechnology and Biosensor Engineering
Molecular Diagnostics
Analytical Chemistry

Background:

Colorimetric aptamer-based sensors offer potential for on-site detection but often lack naked-eye sensitivity.
Existing sensors struggle with efficient target recognition and signal reporting for visual detection.
There is a need for improved sensor platforms for rapid, instrument-free small molecule detection.

Purpose of the Study:

To develop a generalizable strategy for engineering highly sensitive, visual colorimetric sensors.
To create novel multimodule split DNA constructs (CBSAzymes) for enhanced analyte detection.
To demonstrate the application of CBSAzymes for detecting small molecules like illicit drugs.

Main Methods:

Engineered novel multimodule split DNA constructs termed "CBSAzymes".
Utilized a cooperative binding split aptamer (CBSA) for target recognition.
Employed a highly active split DNAzyme for signal reporting, catalyzing a colorimetric reaction.

Main Results:

CBSAzymes assemble upon target binding to form a complex that catalyzes a reaction, producing a visible dark green color within 5 minutes.
Achieved naked-eye detection of cocaine at concentrations as low as 10 μM using a cocaine-binding CBSAzyme.
Developed CBSAzymes for visual detection of MDPV and 10 other synthetic cathinones in biological samples at low micromolar concentrations.

Conclusions:

CBSAzymes represent a generalizable and straightforward strategy for engineering advanced aptamer-DNAzyme biosensors.
These assays enable rapid, sensitive, and instrument-free visual detection of various small molecules.
CBSAzyme-based assays show significant potential for on-site detection of illicit drugs, biomarkers, and toxins.