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Microfluidic bead-based biosensor: Ultrasensitive ctDNA detection based on duplex-functional split-DNAzyme and
1Hunan Provincial Key Laboratory of Environmental Catalysis and Waste Recycling, College of Materials and Chemical Engineering, Hunan Institute of Engineering, Xiangtan, 411104, China.
Analytical Biochemistry
|January 6, 2024
Summary
This study presents a novel ultrasensitive biosensor for detecting Epstein-Barr virus (EBV) DNA, a key biomarker for nasopharyngeal carcinoma. The method uses a cascade signal amplification strategy for early cancer detection using circulating tumor DNA (ctDNA).
Area of Science:
- Biomedical Engineering
- Molecular Diagnostics
- Cancer Biomarkers
Background:
- Circulating tumor DNA (ctDNA) is vital for noninvasive cancer monitoring.
- Developing ultrasensitive and selective assays for ctDNA is crucial for early diagnosis and treatment.
- Nasopharyngeal carcinoma-associated Epstein-Barr virus (EBV) DNA is a significant biomarker.
Purpose of the Study:
- To develop an ultrasensitive and specific detection method for EBV DNA using a microfluidic microbead array chip.
- To utilize a cascade signal amplification strategy involving duplex-functional split-DNAzyme and dendritic probes.
- To establish a promising platform for early-stage ctDNA analysis.
Main Methods:
- A cascade signal amplification strategy was employed using duplex-functional split-DNAzyme and dendritic probes.
- Detection was performed on microfluidic microbead array chips for EBV DNA.
- The system leveraged Pb2+-assisted DNAzyme activity, target recycling, and dendritic enzyme-free signal amplification.
Main Results:
- An excellent detection limit of 0.36 fM and a wide linear range of 1 fM–103 fM were achieved.
- The assay demonstrated high sensitivity and specificity for EBV DNA detection.
- Simulated sample analysis showed recovery rates from 97.2% to 108.1% with low relative standard deviation (3.3%–5.9%).
Conclusions:
- The developed microfluidic biosensor is a highly sensitive and anti-interference system for ctDNA analysis.
- Minimal reagent volumes are required, making the platform cost-effective.
- This technology holds promise for detecting ctDNA at the earliest stages of cancer incidence.
