Method for microRNA isolation from clinical serum samples.
Pubmed ID: 22982505
Pubmed Central ID: PMC3481852
Journal: Analytical biochemistry
Publication Date: 12/01/2012
Affiliation: Benaroya Research Institute and Center for Liver Disease, Digestive Disease Institute, Virginia Mason Medical Center, Seattle, WA 98101, USA. firstname.lastname@example.org
MeSH Terms: Humans, Animals, Double-Blind Method, RNA, Caenorhabditis elegans, MicroRNAs, Real-Time Polymerase Chain Reaction, Biomarkers
Grants: 1R21HL112678, 3R01DK056924-08S1, 5K24DK002957, K24 DK002957, R01 DK056924, R21 HL112678
Authors: Li Y, Kowdley KV
Cite As: Li Y, Kowdley KV. Method for microRNA isolation from clinical serum samples. Anal Biochem 2012 Dec 1;431(1):69-75. Epub 2012 Sep 11.
MicroRNAs are a group of intracellular noncoding RNA molecules that have been implicated in a variety of human diseases. Because of their high stability in blood, microRNAs released into circulation could be potentially utilized as noninvasive biomarkers for diagnosis or prognosis. Current microRNA isolation protocols are specifically designed for solid tissues and are impractical for biomarker development utilizing small-volume serum samples on a large scale. Thus, a protocol for microRNA isolation from serum is needed to accommodate these conditions in biomarker development. To establish such a protocol, we developed a simplified approach to normalize sample input by using single synthetic spike-in microRNA. We evaluated three commonly used commercial microRNA isolation kits for the best performance by comparing RNA quality and yield. The manufacturer's protocol was further modified to improve the microRNA yield from 200μl of human serum. MicroRNAs isolated from a large set of clinical serum samples were tested on the miRCURY LNA real-time PCR panel and confirmed to be suitable for high-throughput microRNA profiling. In conclusion, we have established a proven method for microRNA isolation from clinical serum samples suitable for microRNA biomarker development.