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Related Experiment Video

Updated: May 23, 2026

Quantitative Real-Time Polymerase Chain Reaction Evaluation of MicroRNA Expression in Kidney and Serum of Mice with Age-Dependent Renal Impairment
06:48

Quantitative Real-Time Polymerase Chain Reaction Evaluation of MicroRNA Expression in Kidney and Serum of Mice with Age-Dependent Renal Impairment

Published on: April 29, 2022

Comprehensive evaluation of reference miRNA for extracellular miRNA quantification in CKD.

Anvesha Srivastava1, Mark Maienschein-Cline2, Ana Pabalan1

  • 1Division of Renal Diseases and Hypertension, George Washington University, Washington DC, USA.

Methods (San Diego, Calif.)
|May 21, 2026
PubMed
Summary

This study identified hsa-miR-24-3p as the most stable endogenous reference microRNA (miRNA) for normalizing extracellular miRNA quantification in plasma. Urinary miRNAs require further optimization for reliable normalization in chronic kidney disease research.

Keywords:
Circulatory miRNAEndogenous controlPlasmaRT-qPCRUrine

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Last Updated: May 23, 2026

Quantitative Real-Time Polymerase Chain Reaction Evaluation of MicroRNA Expression in Kidney and Serum of Mice with Age-Dependent Renal Impairment
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A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model
07:01

A Quantitative Detection Method for MicroRNAs in the Kidney of an Ischemic Kidney Injury Mouse Model

Published on: September 11, 2020

Area of Science:

  • Biomolecular analysis
  • Molecular diagnostics
  • Biomarker discovery

Background:

  • Extracellular microRNAs (miRNAs) are promising biomarkers and therapeutic targets.
  • Reliable quantification of extracellular miRNAs using RT-qPCR necessitates appropriate normalization.
  • A universal endogenous reference miRNA for extracellular analysis has not been established.

Purpose of the Study:

  • To identify optimal endogenous control miRNAs for extracellular miRNA quantification.
  • To assess miRNA stability in plasma and urine samples from chronic kidney disease patients.

Main Methods:

  • Next-generation sequencing identified candidate reference miRNAs in plasma and urine.
  • NormFinder algorithm and complementary stability metrics prioritized candidates.
  • RT-qPCR validated top candidate miRNA stability in independent cohorts.

Main Results:

  • Hsa-miR-24-3p demonstrated superior expression stability and detectability in plasma (97%), establishing it as a robust endogenous reference.
  • Hsa-miR-126-3p and hsa-miR-30e-5p also showed high stability in plasma; dual normalization with hsa-miR-30e-5p improved stability.
  • Urinary reference miRNAs exhibited low detectability and insufficient stability for routine normalization without optimization.

Conclusions:

  • Hsa-miR-24-3p is a reliable endogenous control for plasma miRNA normalization in chronic kidney disease.
  • Urinary miRNA quantification requires context-specific validation and technical optimization for accurate results.
  • This study provides crucial guidance for accurate and reproducible extracellular miRNA analysis.