Regular article
Plasma Components Affect Accuracy of Circulating Cancer-Related MicroRNA Quantitation

https://doi.org/10.1016/j.jmoldx.2011.09.002Get rights and content
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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.

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Supported by the US Army Medical Research and Materiel Command under award W81XWH-08-1-0641, research grant 189903 from the American Cancer Society of Illinois (D.M.D.), NIH grant R01 CA119903 (J.N.M.), and grant T32-AI007392/AI/NIAID NIH HHS (S.D.L.).

Opinions, conclusions, interpretations, and recommendations are those of the authors and are not necessarily endorsed by the US Army.

D.M.D., D.J.K., and S.L. conceived and designed the experiments; D.J.K., S.L., and J.C.P. performed the experiments; D.M.D., D.J.K., S.L., M.L.H., and J.C.P. analyzed the data; E.N., J.K.-K., J.N.M., K.B., and M.L.H. contributed reagents/materials/analysis tools; and D.J.K. and D.M.D. wrote the article.

Supplemental material for this manuscript can be found at http://jmd.amjpathol.org or at doi: 10.1016/j.jmoldx.2011.09.002.

Current addresses: D.J.K., Pathology Laboratory of Shin, Daegu, South Korea; J.C.P., Department of Obstetrics and Gynecology, School of Medicine, Keimyung University, Daegu, South Korea.