title: Standardizing analysis of circulating microRNA: clinical and biological relevance.
abstract: Circulating microRNAs (c-miRNAs) provide a new dimension as clinical biomarkers for disease diagnosis, progression, and response to treatment. However, the discovery of individual miRNAs from biofluids that reliably reflect disease states is in its infancy. The highly variable nature of published studies exemplifies a need to standardize the analysis of miRNA in circulation. Here, we show that differential sample handling of serum leads to inconsistent and incomparable results. We present a standardized method of RNA isolation from serum that eliminates multiple freeze/thaw cycles, provides at least three normalization mechanisms, and can be utilized in studies that compare both archived and prospectively collected samples. It is anticipated that serum processed as described here can be profiled, either globally or on a gene by gene basis, for c-miRNAs and other non-coding RNA in the circulation to reveal novel, clinically relevant epigenetic signatures for a wide range of diseases.
authors.affiliation: Vermont Cancer Center for Basic and Translational Research, University of Vermont College of Medicine, Burlington, Vermont, 05405; Department of Biochemistry, University of Vermont College of Medicine, Burlington, Vermont, 05405.
authors.forename: Nicholas H
authors.initials: NH
authors.lastname: Farina
authors.forename: Marie E
authors.initials: ME
authors.lastname: Wood
authors.forename: Scott D
authors.initials: SD
authors.lastname: Perrapato
authors.forename: Christopher S
authors.initials: CS
authors.lastname: Francklyn
authors.forename: Gary S
authors.initials: GS
authors.lastname: Stein
authors.forename: Janet L
authors.initials: JL
authors.lastname: Stein
authors.forename: Jane B
authors.initials: JB
authors.lastname: Lian
mesh_terms: D000818:Animals; D015415:Biomarkers / Q000097:blood*; D020869:Gene Expression Profiling; D006801:Humans; D008297:Male; D051379:Mice; D035683:MicroRNAs / Q000097:blood* / Q000302:isolation & purification; D011471:Prostatic Neoplasms / Q000097:blood* / Q000175:diagnosis / Q000473:pathology; D012015:Reference Standards
chemical_list: D015415:Biomarkers; D035683:MicroRNAs
keywords: BIOMARKER; CIRCULATING microRNA; CLINICAL SAMPLE; PLASMA; RNA ISOLATION; SERUM; c-miRNA; miRNA; qPCR