Related Experiment Video
Updated: Jan 5, 2026
MicroRNA Based Liquid Biopsy: The Experience of the Plasma miRNA Signature Classifier MSC for Lung Cancer Screening
Published on: October 26, 2017
Plasma microRNA detection standardization test
Xiaomin Feng1, Yuwei Liu1, Nan Wan2
1Dalian Medical University, Dalian, Liaoning, China.
Background:
MicroRNAs (miRNAs) can be used for early diagnosis of myocardial infarction. However, due to a lack of standardized operating procedures, their value for clinical application is low.
Methods:
Detection of plasma miRNAs was optimized by analyzing factors influencing miRNA variance and myocardial infarction risk scores during analysis (extraction, reverse transcription, and real-time PCR) and pre-analysis (dietary status, anticoagulants, storage conditions, and hemolysis).
Results:
Regarding variable factors during analysis, the centrifugal column method was superior to Trizol LS reagent when extracting miRNA from plasma. Recovery rate was highest with plasma volumes of 200 and 300 µL. During analysis, the main source of miRNA detection inaccuracy was derived from RNA extraction (mainly organic extraction), and not reverse transcription or PCR. MiRNA variance could be reduced by use of an internal reference. During analysis, 95% of risk score variation fluctuated within a range of 6.267. The variable factors pre-analysis mainly involved dietary status, anticoagulant selection, and storage conditions. Hemolysis positively correlated with miRNA levels, but there was no significant change in risk score after internal reference calibration.
Conclusion:
Preliminary standardization for miRNA detection provides a reference for clinical blood testing of miRNAs.
Insights
Standardizing microRNA (miRNA) detection improves accuracy for diagnosing myocardial infarction. Optimizing extraction and pre-analysis factors enhances clinical reliability of plasma miRNA testing.
Area of Science:
- Biochemistry
- Molecular Biology
- Clinical Diagnostics
Background:
- MicroRNAs (miRNAs) show promise for early myocardial infarction diagnosis.
- Current clinical application is limited by a lack of standardized detection procedures.
Purpose of the Study:
- To optimize plasma miRNA detection methods.
- To identify factors affecting miRNA variance and myocardial infarction risk scores.
Main Methods:
- Analyzed factors influencing miRNA detection during extraction, reverse transcription, and real-time PCR.
- Evaluated pre-analysis variables including dietary status, anticoagulants, storage, and hemolysis.
- Compared centrifugal column method with Trizol LS reagent for RNA extraction.
Main Results:
- Centrifugal column method outperformed Trizol LS for plasma miRNA extraction.
- Optimal plasma volume for recovery was 200-300 µL.
- RNA extraction, particularly organic extraction, was the primary source of inaccuracy; internal references reduced variance.
Conclusions:
- Preliminary standardization of miRNA detection offers a reference for clinical blood testing.
- Optimized methods enhance the reliability of miRNA-based diagnostics for myocardial infarction.
