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Highly efficient and automated isolation technology for extracellular vesicles microRNA
Kaili Di1, Boyue Fan2, Xinrui Gu1
1Department of Laboratory Medicine, Affiliated Drum Tower Hospital, Medical School of Nanjing University, Nanjing, China.
Abstract:
MicroRNA (miRNA) in extracellular vesicles (EVs) has great potential to be a promising marker in liquid biopsy. However, the present EV isolation methods, such as ultracentrifugation, have complicated and long-time operation, which impedes research on EV miRNA. The downstream complex miRNA extraction process will also significantly increase the detection cycle and loss. We first established a simple automated technique to efficiently extract target miRNAs in EVs from plasma based on Fe3O4@TiO2 beads with high affinity and capture efficiency. We combined a heat-lysis method for quick and simple EV miRNA extraction and detection. The results indicated that our method has more RNA yield than TRIzol or a commercial kit and could complete EV enrichment and miRNA extraction in 30 min. Through the detection of miRNA-21, healthy people and lung cancer patients were distinguished, which verified the possibility of the application in clinical detection. The automated isolation technology for EV miRNA has good repeatability and high throughput, with great application potential in clinical diagnosis.
Insights
This study introduces a rapid, automated method using Fe3O4@TiO2 beads for isolating microRNA (miRNA) from extracellular vesicles (EVs). This technique simplifies liquid biopsy analysis for potential lung cancer detection.
Area of Science:
- Biochemistry
- Molecular Biology
- Nanotechnology
Background:
- Extracellular vesicles (EVs) contain microRNAs (miRNAs) with diagnostic potential for liquid biopsies.
- Current EV isolation and miRNA extraction methods are time-consuming and complex, hindering clinical applications.
- Efficient and rapid isolation of EV-derived miRNAs is crucial for advancing liquid biopsy technologies.
Purpose of the Study:
- To develop a simple, automated technique for efficient extraction of target miRNAs from plasma-derived EVs.
- To combine heat-lysis for rapid EV miRNA extraction and detection.
- To assess the clinical applicability of the developed method for distinguishing lung cancer patients from healthy individuals.
Main Methods:
- Utilized Fe3O4@TiO2 magnetic beads for high-affinity capture and isolation of EVs from plasma.
- Integrated a heat-lysis method for rapid and straightforward extraction of miRNAs from isolated EVs.
- Employed miRNA-21 as a biomarker to differentiate between healthy individuals and lung cancer patients.
Main Results:
- The automated method demonstrated higher RNA yield compared to TRIzol and a commercial kit.
- EV enrichment and miRNA extraction were completed within 30 minutes.
- Successful differentiation between healthy individuals and lung cancer patients using miRNA-21 detection, validating clinical potential.
Conclusions:
- The developed automated Fe3O4@TiO2 bead-based technique offers an efficient and rapid solution for EV miRNA isolation and detection.
- This method significantly reduces processing time and complexity compared to traditional techniques.
- The technology exhibits good repeatability and high throughput, showing great promise for clinical diagnosis and liquid biopsy applications.
