Document Detail

Sensitive PCR-based quantitation of cell-free circulating microRNAs.
MedLine Citation:
PMID:  22884953     Owner:  NLM     Status:  MEDLINE    
Cell-free microRNAs (miRNAs) that circulate in the blood are promising surrogate biomarkers of disease and physiological processes. The ease of quantifying specific miRNA species using made-to-order approaches based on Taq-polymerase has led to numerous studies that have identified changes in the abundance of circulating cell-free miRNA species that correlate with pathology or other events. The growing interest in developing miRNAs as blood biomarkers necessitates the careful consideration of the unique properties of such body fluids that can make the reproducible and quantitative assessment of RNA abundance challenging. For example, enzymes involved in the amplification and analysis of RNA can be affected by blood components that copurify with miRNA. Thus, if miRNAs are to be effectively utilized as biomarkers, it is important to establish standardized protocols for blood collection and miRNA analysis to ensure accurate quantitation. Here we outline several considerations, including the type of collection tube used in sampling, the influence of added anticoagulants and stabilizers, sample processing, enrichment of vesicular and other miRNA species, RNA extraction approaches and enzyme selection, that affect quantitation of miRNA isolated from plasma and should be considered in order to achieve reproducible, sensitive and accurate quantitation.
Michelle L Hastings; Jaime Palma; Dominik M Duelli
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Publication Detail:
Type:  Journal Article; Research Support, N.I.H., Extramural; Research Support, Non-U.S. Gov't; Research Support, U.S. Gov't, Non-P.H.S.     Date:  2012-08-03
Journal Detail:
Title:  Methods (San Diego, Calif.)     Volume:  58     ISSN:  1095-9130     ISO Abbreviation:  Methods     Publication Date:  2012 Oct 
Date Detail:
Created Date:  2012-11-26     Completed Date:  2013-05-24     Revised Date:  2013-10-17    
Medline Journal Info:
Nlm Unique ID:  9426302     Medline TA:  Methods     Country:  United States    
Other Details:
Languages:  eng     Pagination:  144-50     Citation Subset:  IM    
Copyright Information:
Copyright © 2012 Elsevier Inc. All rights reserved.
Department of Cell Biology and Anatomy, Rosalind Franklin University of Medicine and Science, North Chicago, IL 60064, USA.
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MeSH Terms
Biological Markers / blood*
Cell-Free System
MicroRNAs* / blood,  isolation & purification
Polymerase Chain Reaction / methods*
RNA* / blood,  genetics,  isolation & purification
Grant Support
Reg. No./Substance:
0/Biological Markers; 0/MicroRNAs; 63231-63-0/RNA

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