Jack W Sharkey1, Daniel J Antoine, B Kevin Park. 1. MRC Centre for Drug Safety Science, Department of Molecular & Clinical Pharmacology, Institute of Translational Medicine, University of Liverpool, United Kingdom.
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.
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. elegansLin-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: Joanna I Clarke; Shiva Seyed Forootan; Jonathan D Lea; Lawrence S Howell; Josep Monne Rodriguez; Anja Kipar; Christopher E Goldring; B Kevin Park; Ian M Copple; Daniel J Antoine Journal: Toxicol Res (Camb) Date: 2017-04-11 Impact factor: 3.524
Authors: Matthew B Hudson; Myra E Woodworth-Hobbs; Bin Zheng; Jill A Rahnert; Mitsi A Blount; Jennifer L Gooch; Charles D Searles; S Russ Price Journal: Am J Physiol Cell Physiol Date: 2013-12-11 Impact factor: 4.249
Authors: Matthew B Hudson; Jill A Rahnert; Bin Zheng; Myra E Woodworth-Hobbs; Harold A Franch; S Russ Price Journal: Am J Physiol Cell Physiol Date: 2014-05-28 Impact factor: 4.249