title: RNA Detection in Urine: From RNA Extraction to Good Normalizer Molecules.
abstract: RNA detection in liquid urine biopsy specimens could be an optimal method for noninvasive diagnostic and prognostic procedures in urologic disorders; however, there are no standardized procedures for implementing it in the clinic. We present a systematic evaluation of the best storage conditions and purification methods using four commercially available extraction kits to purify RNA from void urine. We measured different RNA molecules to select good and stable biomarkers and normalizers for analyses in liquid urine biopsy specimens. We have established a new combined procedure for RNA isolation from urine and found good performance in 25 urine samples from healthy volunteers of both sexes. Associations were tested using the t-test for paired samples, and miRNA specimens were selected as the more stable molecules. Stability analysis was performed, and we found miR193a and miR448 as the best normalizers to be used in this biofluid. This is a highly reproducible method that could be used to evaluate urine samples for diagnostic and prognostic purposes.
authors.affiliation: Molecular Oncology Unit, Research Center for Energy, Environment, and Technology [CIEMAT (ed70A)], Madrid; and the Uro-Oncology Unit, University Hospital 12 de Octubre, Madrid, Spain.
authors.forename: Mónica
authors.initials: M
authors.lastname: Martínez-Fernández
authors.affiliation: Molecular Oncology Unit, Research Center for Energy, Environment, and Technology [CIEMAT (ed70A)], Madrid; and the Uro-Oncology Unit, University Hospital 12 de Octubre, Madrid, Spain. Electronic address: jesusm.paramio@ciemat.es.
authors.forename: Jesús M
authors.initials: JM
authors.lastname: Paramio
authors.affiliation: Molecular Oncology Unit, Research Center for Energy, Environment, and Technology [CIEMAT (ed70A)], Madrid; and the Uro-Oncology Unit, University Hospital 12 de Octubre, Madrid, Spain. Electronic address: marta.duenas@ciemat.es.
authors.forename: Marta
authors.initials: M
authors.lastname: Dueñas
mesh_terms: D000328:Adult; D000368:Aged; D015415:Biomarkers / Q000652:urine; D005260:Female; D006801:Humans; D008297:Male; D035683:MicroRNAs / Q000032:analysis / Q000652:urine*; D008875:Middle Aged; D025202:Molecular Diagnostic Techniques / Q000379:methods*; D055815:Young Adult
chemical_list: D015415:Biomarkers; C546818:MIRN193 microRNA, human; C557400:MIRN448 microRNA, human; D035683:MicroRNAs