Direct screening of plasma circulating microRNAs.

Area of Science:

• Biochemistry
• Molecular Biology
• Genomics

Background:

• Circulating microRNAs (miRNAs) are promising biomarkers for disease detection.
• RNA-Sequencing is a standard method for miRNA quantification but involves extensive sample preparation.
• Streamlining miRNA analysis is crucial for efficient biomarker discovery.

Purpose of the Study:

• To evaluate the HTG EdgeSeq technology for circulating miRNA profiling in human plasma.
• To assess the reliability and efficiency of HTG EdgeSeq by comparing it with established methods.
• To determine the potential of HTG EdgeSeq for large-scale miRNA biomarker discovery.

Main Methods:

• HTG EdgeSeq technology was applied to human plasma specimens, omitting the RNA extraction step.
• Quantitative Polymerase Chain Reaction (qPCR) and digital Polymerase Chain Reaction (dPCR) were used for comparison.
• Correlation analysis (Pearson's coefficient) was performed to compare results between methods.

Main Results:

• HTG EdgeSeq significantly reduced hands-on steps, preparation time, and costs compared to traditional RNA-Sequencing.
• A strong positive correlation was observed between HTG EdgeSeq and qPCR (Pearson's coefficient = 0.93).
• A strong positive correlation was also observed between HTG EdgeSeq and dPCR (Pearson's coefficient = 0.94).

Conclusions:

• HTG EdgeSeq technology is a reliable method for analyzing circulating miRNAs in human plasma without prior RNA extraction.
• The technology enables simultaneous screening of over 2,000 miRNAs, offering high throughput.
• HTG EdgeSeq is suitable for biomarker discovery in large patient cohorts due to its efficiency and reliability.