title: Validation of the isolation and quantification of kidney enriched miRNAs for use as biomarkers.
abstract: BACKGROUND
  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.
authors.affiliation: MRC Centre for Drug Safety Science, Department of Molecular & Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, United Kingdom.
authors.forename: Jack W
authors.initials: JW
authors.lastname: Sharkey
authors.forename: Daniel J
authors.initials: DJ
authors.lastname: Antoine
authors.forename: B Kevin
authors.initials: BK
authors.lastname: Park
mesh_terms: D001483:Base Sequence; D015415:Biomarkers / Q000032:analysis*; D017931:DNA Primers; D006801:Humans; D007668:Kidney / Q000378:metabolism*; D035683:MicroRNAs / Q000032:analysis*; D016133:Polymerase Chain Reaction
chemical_list: D015415:Biomarkers; D017931:DNA Primers; D035683:MicroRNAs