Assessment of Urinary MicroRNAs by Quantitative Polymerase Chain Reaction in Diabetic Nephropathy Patients

  • Lucy Jade Newbury
  • Alexa Wonnacott
  • Kate Simpson
  • Timothy Bowen
  • Donald FraserEmail author
Part of the Methods in Molecular Biology book series (MIMB, volume 2067)


Urinary microRNAs show promise as noninvasive biomarkers in renal disease. Here, we describe a detailed protocol for the column-based extraction and quantification of miRNAs by reverse-transcription quantitative polymerase chain reaction (RT-qPCR) from urine samples.

Key words

MicroRNAs Urine Biomarker Diabetic nephropathy Chronic kidney disease RT-qPCR 


  1. 1.
    Jha V, Garcia-Garcia G, Iseki K, Li Z, Naicker S, Plattner B, Saran R, Wang AY, Yang CW (2013) Chronic kidney disease: global dimension and perspectives. Lancet 382(9888):260–272. Elsevier Ltd, England. Scholar
  2. 2.
    Cowie CC, Port FK, Wolfe RA, Savage PJ, Moll PP, Hawthorne VM (1989) Disparities in incidence of diabetic end-stage renal disease according to race and type of diabetes. N Engl J Med 321(16):1074–1079. Scholar
  3. 3.
    Regele F, Jelencsics K, Shiffman D, Pare G, McQueen MJ, Mann JF, Oberbauer R (2015) Genome-wide studies to identify risk factors for kidney disease with a focus on patients with diabetes. Nephrol Dial Transplant 30(Suppl 4):iv26–iv34. The Author 2015. Published by Oxford University Press on behalf of ERA-EDTA, England. Scholar
  4. 4.
    Parving HH, Persson F, Rossing P (2015) Microalbuminuria: a parameter that has changed diabetes care. Diabetes Res Clin Pract 107(1):1–8. Elsevier Ireland Ltd, Ireland. Scholar
  5. 5.
    Landgraf P, Rusu M, Sheridan R, Sewer A, Iovino N, Aravin A, Pfeffer S, Rice A, Kamphorst AO, Landthaler M, Lin C, Socci ND, Hermida L, Fulci V, Chiaretti S, Foa R, Schliwka J, Fuchs U, Novosel A, Muller RU, Schermer B, Bissels U, Inman J, Phan Q, Chien M, Weir DB, Choksi R, De Vita G, Frezzetti D, Trompeter HI, Hornung V, Teng G, Hartmann G, Palkovits M, Di Lauro R, Wernet P, Macino G, Rogler CE, Nagle JW, Ju J, Papavasiliou FN, Benzing T, Lichter P, Tam W, Brownstein MJ, Bosio A, Borkhardt A, Russo JJ, Sander C, Zavolan M, Tuschl T (2007) A mammalian microRNA expression atlas based on small RNA library sequencing. Cell 129(7):1401–1414CrossRefGoogle Scholar
  6. 6.
    Arroyo JD, Chevillet JR, Kroh EM, Ruf IK, Pritchard CC, Gibson DF, Mitchell PS, Bennett CF, Pogosova-Agadjanyan EL, Stirewalt DL, Tait JF, Tewari M (2011) Argonaute 2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc Natl Acad Sci U S A 108(12):5003–5008CrossRefGoogle Scholar
  7. 7.
    Beltrami C, Clayton A, Newbury LJ, Corish P, Jenkins RH, Phillips AO, Fraser DJ, Bowen T (2015) Stabilization of urinary microRNAs by association with exosomes and argonaute 2 protein. Noncoding RNA 1(2):151–166. Scholar
  8. 8.
    Smith DA, Newbury LJ, Drago G, Bowen T, Redman JE (2017) Electrochemical detection of urinary microRNAs via sulfonamide-bound antisense hybridisation. Sensors Actuators B Chem 253. (Suppl C:335–341. Scholar
  9. 9.
    Trionfini P, Benigni A, Remuzzi G (2015) MicroRNAs in kidney physiology and disease. Nat Rev Nephrol 11(1):23–33. England. Scholar
  10. 10.
    Simpson K, Wonnacott A, Fraser DJ, Bowen T (2016) MicroRNAs in diabetic nephropathy: from biomarkers to therapy. Curr Diab Rep 16(3):35. Scholar
  11. 11.
    Krupa A, Jenkins R, Luo DD, Lewis A, Phillips A, Fraser D (2010) Loss of MicroRNA-192 promotes fibrogenesis in diabetic nephropathy. J Am Soc Nephrol 21(3):438–447. Scholar
  12. 12.
    Heinzel A, Muhlberger I, Stelzer G, Lancet D, Oberbauer R, Martin M, Perco P (2015) Molecular disease presentation in diabetic nephropathy. Nephrol Dial Transplant 30(Suppl 4):iv17–iv25. The Author 2015. Published by Oxford University Press on behalf of ERA-EDTA, England. Scholar
  13. 13.
    Lopez-Anton M, Lambie M, Lopez-Cabrera M, Schmitt CP, Ruiz-Carpio V, Bartosova M, Schaefer B, Davies S, Stone T, Jenkins R, Taylor PR, Topley N, Bowen T, Fraser D (2017) miR-21 promotes fibrogenesis in peritoneal dialysis. Am J Pathol 187(7):1537–1550. Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Lucy Jade Newbury
    • 1
  • Alexa Wonnacott
    • 1
  • Kate Simpson
    • 1
  • Timothy Bowen
    • 1
  • Donald Fraser
    • 1
    Email author
  1. 1.Wales Kidney Research UnitHeath Park CampusCardiffUK

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