NGS and qPCR analysis of RNA and microRNA in biofluids and exosomes

Michael Thorsen 1,
1Exiqon, Denmark; 

Abstract
Biofluids contain a multitude of RNA molecules some of which may serve as biomarkers for altered conditions in the body and be applied in diagnosis of disease. Many biofluids are easily collected and liquid biopsies hold the promise of developing non- or mini- mal invasive diagnostic tests for screening and monitoring diseases such as cancer. Though rather unstable by nature, RNA in biofluids or a fraction of these appear relatively stable, possibly because they are protected from degradation within exosomes or within protein aggregates.
We are developing technologies and offer services to analyze RNA in biofluids which is challenged by the low content of RNA and the presence of substances that interfere with the analyses. Exo- somes are of particular interest, as the RNA cargo that these vesicles carry are derived from cells throughout the body and in partic- ular the cells in contact with the biofluid. Currently, we are able to robustly analyze RNA including microRNA in different biofluids and exosomes using both next generation sequencing and LNATM enhanced qPCR. Data will be presented showing how miRCURY LNATM Universal RT microRNA PCR may be applied successfully to validate NGS data. We will also share data on applying qPCR analysis of urinary exosomes to discover microRNA biomarkers for prostate cancer and how a three-microRNA signature now has been validated in independent cohorts.
http://dx.doi.org/10.1016/j.bdq.2017.02.025


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