BIOMARKERS FROM EXTRACELLULAR VESICLES IN URINE

A Robust, Automated Strategy for Biomarker Discovery from Extracellular Vesicles in Urine

A recent study by Anson Ku and others from Lund University, “A urinary extracellular vesicle microRNA biomarker discovery pipeline; from automated extracellular vesicle enrichment by acoustic trapping to microRNA sequencing”, investigated a comprehensive biomarker discovery pipeline for miRNA derived from both small and large urinary EVs.

Biomarker Discovery Pipeline

The biomarker discover pipeline consists of:

  • Enrichment of urinary EVs using an acoustic trap or ultracentrifugation
  • RNase A treatment to eliminate contaminating RNA molecules
  • Total RNA isolation
  • Small RNA library preparation
  • Illumina® sequencing

Low Input miRNA Library Prep

Libraries were constructed from two commercially available, low-input compatible, small RNA library kits, the NEXTFLEX® Small RNA-seq kit and the CATS small RNA library (Diagenode, USA), using an estimated 130 pg of input total RNA isolated form 1.7 mL of urine. The NEXTFLEX® Small RNA-seq kit because the NEXTFLEX® libraries resulted in more mappable reads compared to the Diagenode® CATS libraries which had a predominant fraction of rRNA reads. Additionally, the NEXTFLEX® Small RNA-seq kit was the only kit able to successfully generate libraries from all the low-input samples.

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Read the research:

Ku, A., Ravi, N., Yang, M., Evander, M., Laurell, T., Lilja, H., & Ceder, Y. (2019). A urinary extracellular vesicle microRNA biomarker discovery pipeline; from automated extracellular vesicle enrichment by acoustic trapping to microRNA sequencing. Plos One, 14(10). doi: 10.1371/journal.pone.0224604.

Learn more about the NEXTFLEX® Small RNA-seq kit and improve your miRNA biomarker pipeline.

For research use only. Not for use in diagnostic procedures.