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Detection of a Circulating MicroRNA Custom Panel in Patients with Metastatic Colorectal Cancer

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Preanalytic Methodological Considerations and Sample Quality Control of Circulating miRNAs.

Suit-Fong Chan¹, He Cheng¹, Karen Kai-Rui Goh¹

¹Research and Development Lab, MiRXES Lab Pte. Ltd., Singapore.

The Journal of Molecular Diagnostics : JMD

|April 8, 2023

Summary

A new 7-microRNA (miRNA) panel aids in controlling sample quality for disease diagnosis. This quality control ensures more reliable detection of circulating miRNAs, crucial for biomarker development.

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Area of Science:

Biochemistry
Molecular Biology
Biomarker Discovery

Background:

Circulating microRNAs (miRNAs) show promise as biomarkers for disease diagnosis and prognosis.
Accurate miRNA quantification relies heavily on pre-analytical sample handling and quality control.
Standardized protocols are needed to mitigate confounding factors affecting miRNA profiles.

Purpose of the Study:

To establish a quality control (QC) panel for circulating miRNA detection.
To identify miRNAs affected by pre-analytical variables like hemolysis and platelet contamination.
To optimize blood sample processing for reliable miRNA analysis.

Main Methods:

Quantitative reverse transcription-polymerase chain reaction (RT-PCR) was used to profile 356 miRNAs.
Analysis included various blood sample types and processing protocols.
Correlation analysis identified miRNAs sensitive to confounding factors and stability issues.

Main Results:

A 7-miRNA panel was developed to assess sample quality, specifically for hemolysis and platelet contamination.
The study identified confounding impacts from blood collection tube size, centrifugation, and storage conditions.
A dual-spin workflow was established for optimal sample processing, and miRNA stability was assessed under various conditions.

Conclusions:

The developed 7-miRNA QC panel effectively assesses sample integrity for circulating miRNA analysis.
Incorporating stability-related miRNAs enhances the QC panel's robustness.
This QC panel supports more reliable and reproducible detection of circulating miRNAs as disease biomarkers.