RNA Detection in Urine: From RNA Extraction to Good Normalizer Molecules

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.