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Sensitive detection of microRNA using a label-free copper nanoparticle system with polymerase-based signal
Jiangnan Lu1, Shuping Liang1, Li Tan1
1Key Laboratory for the Chemistry and Molecular Engineering of Medicinal Resources (Ministry of Education of China), School of Chemistry and Pharmaceutical Science of Guangxi Normal University, Guanxi, Guilin, 541004, China.
Analytical and Bioanalytical Chemistry
|August 14, 2020
Summary
We developed a novel biosensor for detecting microRNAs (miRNAs) using DNA-copper nanoparticle complexes. This sensitive method allows for accurate miRNA expression analysis in biological samples.
Area of Science:
- Biomedical Engineering
- Nanotechnology
- Molecular Biology
Background:
- Abnormal microRNA (miRNA) expression is linked to various diseases, necessitating accurate detection methods.
- Current miRNA detection techniques can be complex and lack sensitivity.
- Developing simple, sensitive, and accurate miRNA detection tools is crucial for disease diagnostics.
Purpose of the Study:
- To develop a novel, sensitive, and simple biosensor for detecting and analyzing microRNA (miRNA) expression.
- To utilize a DNA-copper nanoparticle (CuNP) based system for enhanced fluorescence detection.
- To establish a method applicable for detecting various miRNA biomarkers.
Main Methods:
- A polymeric double-stranded DNA (dsDNA) copper nanoparticle (CuNP) template was synthesized using polymerase.
- The dsDNA template promoted CuNP formation in the presence of Cu2+ and ascorbic acid, leading to fluorescence.
- Fluorescence intensity changes were measured to quantify miRNA levels.
Main Results:
- The biosensor demonstrated sensitive miRNA detection with a limit of detection as low as 17.8 pmol/L.
- A linear detection range for miR-122 was observed from 80.0 pmol/L to 4.50 nmol/L.
- High recovery rates (93.33-102.53%) were achieved in spiked HepG2 cell lysates, indicating accuracy.
Conclusions:
- The developed fluorescent DNA-CuNP biosensor offers a sensitive and accurate method for miRNA expression analysis.
- This label-free, enzyme-assisted method expands the application of fluorescent DNA-CuNPs in biosensing.
- The system's adaptability allows for the detection of any miRNA marker by modifying the target recognition sequence.
