Validation of the isolation and quantification of kidney enriched miRNAs for use as biomarkers

Abstract

Context: MiRNAs have been reported to represent sensitive and translational biomarkers of organ injury

Objective: To validate the methodologies for the isolation and quantification of a miRNAs from none-invasive biofluids

Methods: Commercially available miRNA isolation kits and qPCR was utilised for determination of analyte sensitivity, stability, recovery, and precision

Results: qPCR was highly precise and sensitive for endogenous miRNA quantification (miR-194, LLQ; 0.1 pM). Intra and inter-assay variation remained low (<12%). Variable recovery (54–89%) was controlled for by internal synthetic standards (C. elegans Lin-4)

Conclusions: Translational endogenous miRNAs reflective of renal injury represent stable analytes with a large dynamic range that can be quantified easily in most laboratories

Keywords::

• Nephrotoxicity
• renal
• biomarker
• exosome
• urine