title: Plasma components affect accuracy of circulating cancer-related microRNA quantitation.
abstract: Circulating microRNAs (miRNAs) have emerged as candidate biomarkers of various diseases and conditions including malignancy and pregnancy. This approach requires sensitive and accurate quantitation of miRNA concentrations in body fluids. Herein we report that enzyme-based miRNA quantitation, which is currently the mainstream approach for identifying differences in miRNA abundance among samples, is skewed by endogenous serum factors that co-purify with miRNAs and anticoagulant agents used during collection. Of importance, different miRNAs were affected to varying extent among patient samples. By developing measures to overcome these interfering activities, we increased the accuracy, and improved the sensitivity of miRNA detection up to 30-fold. Overall, the present study outlines key factors that prevent accurate miRNA quantitation in body fluids and provides approaches that enable faithful quantitation of miRNA abundance in body fluids.
authors.affiliation: Department of Pathology, Chicago Medical School, North Chicago, Illinois 60064, USA.
authors.forename: Dong-Ja
authors.initials: DJ
authors.lastname: Kim
authors.forename: Sarah
authors.initials: S
authors.lastname: Linnstaedt
authors.forename: Jaime
authors.initials: J
authors.lastname: Palma
authors.forename: Joon Cheol
authors.initials: JC
authors.lastname: Park
authors.forename: Evangelos
authors.initials: E
authors.lastname: Ntrivalas
authors.forename: Joanne Y H
authors.initials: JY
authors.lastname: Kwak-Kim
authors.forename: Alice
authors.initials: A
authors.lastname: Gilman-Sachs
authors.forename: Kenneth
authors.initials: K
authors.lastname: Beaman
authors.forename: Michelle L
authors.initials: ML
authors.lastname: Hastings
authors.forename: Jeffrey N
authors.initials: JN
authors.lastname: Martin
authors.forename: Dominik M
authors.initials: DM
authors.lastname: Duelli
mesh_terms: D000925:Anticoagulants / Q000737:chemistry; D014408:Biomarkers, Tumor / Q000097:blood*; D002478:Cells, Cultured; D012321:DNA-Directed RNA Polymerases / Q000037:antagonists & inhibitors / Q000737:chemistry; D019763:Heparin Lyase / Q000737:chemistry; D006801:Humans; D035683:MicroRNAs / Q000097:blood*; D009369:Neoplasms / Q000097:blood* / Q000175:diagnosis; D010070:Oxalates / Q000737:chemistry; D010949:Plasma / Q000737:chemistry; D020871:RNA Stability; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods*; D012680:Sensitivity and Specificity; D012969:Sodium Fluoride / Q000737:chemistry; D013048:Specimen Handling
chemical_list: D000925:Anticoagulants; D014408:Biomarkers, Tumor; D012321:DNA-Directed RNA Polymerases; D019763:Heparin Lyase; D035683:MicroRNAs; D010070:Oxalates; D012969:Sodium Fluoride; C505362:MIRN16 microRNA, human; C525067:MIR223, human