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

Biomarkers

MRC Centre for Drug Safety Science, Department of Molecular & Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, United Kingdom.

Published: May 2012

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.

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http://dx.doi.org/10.3109/1354750X.2012.657246DOI Listing

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