REPORTS
opment. If protocols can be developed to enable reverse transcription and PCR directly from patient samples (30), this platform has the potential to make point- of-care diagnostics based upon circu- lating microRNAs a reality.
Author contributions
W.J.A. performed and analyzed the exper- iments and assisted with manuscript preparation. E.B. and M.D. performed experiments with plasma and assisted with manuscript preparation. R.E.H. provided plasma samples and assisted with manuscript preparation. D.A.S. conceived the study, analyzed the data, and wrote the manuscript.
Acknowledgments
This work was supported by grants from the Department of Education and Learning (DEL), Invest Northern Ireland (INI), the European Regional Development Fund, and Fight For Sight.
Competing interests The authors declare no competing interests.
References
1. Laurin, N. and C. Frégeau. 2012. Optimi- zation and validation of a fast amplification protocol for AmpFlSTR® Profiler Plus® for rapid forensic human identification. Forensic Sci. Int. Genet. 6:47-57.
2. Bahlmann, S., S. Hughes-Stamm, and D. Gangitano. 2014. Development and evaluation of a rapid PCR method for the PowerPlex®S5 system for forensic DNA profiling. Leg Med (Tokyo). 16:227-233.
3. Wheeler, E.K., C.A. Hara, J. Frank, J. Deotte, S.B. Hall, W. Benett, C. Spadaccini, and N.R. Beer. 2011. Under-three minute PCR: probing the limits of fast amplification. Analyst. 136:3707-3712.
4. Neuzil, P., C. Zhang, J. Pipper, S. Oh, and L. Zhuo. 2006. Ultra fast miniaturized real-time PCR: 40 cycles in less than six minutes. Nucleic Acids Res. 34:e77.
5. Hashimoto, M., P.-C. Chen, M.W. Mitchell, D.E. Nikitopoulos, S.A. Soper, and M.C. Murphy. 2004. Rapid PCR in a continuous flow device. Lab Chip 4:638-645.
6. Farrar, J.S. and C.T. Wittwer. 2015. Extreme PCR: Efficient and Specific DNA Amplification in 15-60 Seconds. Clin. Chem. 61:145-153.
7. Wittwer, C.T., K.M. Ririe, R.V. Andrew, D.A. David, R.A. Gundry, and U.J. Balis. 1997. The LightCycler: a microvolume multisample fluorimeter with rapid temperature control. Biotechniques 22:176-181.
8. Mitchell, P.S., R.K. Parkin, E.M. Kroh, B.R. Fritz, S.K. Wyman, E.L. Pogosova-
Vol. 58 | No. 5 | 2015
Agadjanyan, A. Peterson, J. Noteboom, et al. 2008. Circulating microRNAs as stable blood-based markers for cancer detection. Proc. Natl. Acad. Sci. USA 105:10513-10518.
9. Chen, X., Y. Ba, L. Ma, X. Cai, Y. Yin, K. Wang, J. Guo, Y. Zhang, et al. 2008. Characterization of microRNAs in serum: a novel class of biomarkers for diagnosis of cancer and other diseases. Cell Res. 18:997- 1006.
10. Creemers, E.E., A.J. Tijsen, and Y.M. Pinto. 2012. Circulating microRNAs: novel biomarkers and extracellular communi- cators in cardiovascular disease? Circ. Res. 110:483-495.
11. Weiland, M., X.-H. Gao, L. Zhou, and Q.-S. Mi. 2012. Small RNAs have a large impact: circulating microRNAs as biomarkers for human diseases. RNA Biol. 9:850-859.
12. Leidinger, P., C. Backes, S. Deutscher, K. Schmitt, S.C. Mueller, K. Frese, J. Haas, K. Ruprecht, et al. 2013. A blood based 12-miRNA signature of Alzheimer disease patients. Genome Biol. 14:R78.
13. Deddens, J.C., J.M. Colijn, M.I.F.J. Oerlemans, G. Pasterkamp, S.A. Chamuleau, P.A. Doevendans, and J.P.G. Sluijter. 2013. Circulating microRNAs as novel biomarkers for the early diagnosis of acute coronary syndrome. J. Cardiovasc. Transl. Res. 6:884-898.
14. Shi, R. and V.L. Chiang. 2005. Facile means for quantifying microRNA expression by real-time PCR. Biotechniques 39:519-525.
15. Levesque-Sergerie, J.-P., M. Duquette, C. Thibault, L. Delbecchi, and N. Bissonnette. 2007. Detection limits of several commercial reverse transcriptase enzymes: impact on the low- and high-abundance transcript levels assessed by quantitative RT-PCR. BMC Mol. Biol. 8:93.
16. Selcuklu, S.D., M.T.A. Donoghue, and C. Spillane. 2009. miR-21 as a key regulator of oncogenic processes. Biochem. Soc. Trans. 37:918-925.
17. Iorio, M.V., M. Ferracin, C.-G. Liu, A. Veronese, R. Spizzo, S. Sabbioni, E. Magri, M. Pedriali, et al. 2005. MicroRNA Gene Expression Deregulation in Human Breast Cancer. Cancer Res. 65:7065-7070.
18. Volinia, S., G.A. Calin, C.-G. Liu, S. Ambs, A. Cimmino, F. Petrocca, R. Visone, M. Iorio, et al. 2006. A microRNA expression signature of human solid tumors defines cancer gene targets. Proc. Natl. Acad. Sci. USA 103:2257-2261.
19. Thum, T., C. Gross, J. Fiedler, T. Fischer, S. Kissler, M. Bussen, P. Galuppo, S. Just, et al. 2008. MicroRNA-21 contributes to myocardial disease by stimulating MAP kinase signal l ing in f ibroblasts. Nature 456:980-984.
20. Asaga, S., C. Kuo, T. Nguyen, M. Terpenning, A.E. Giuliano, and D.S.B. Hoon. 2011. Di rect serum assay for microRNA-21 concentrations in early and advanced breast cancer. Cl in. Chem. 57:84-91.
21. Xu, J., C. Wu, X. Che, L. Wang, D. Yu, T. Zhang, L. Huang, H. Li, et al. 2011. Circulating MicroRNAs, miR-21, miR-122, and miR-223,
in patients with hepatocel- 252
lular carcinoma or chronic hepatitis. Mol. Carcinog. 50:136-142.
22. Zhang, J., Y. Yang, T. Yang, Y. Liu, A. Li, S. Fu, M. Wu, Z. Pan, and W. Zhou. 2010. microRNA-22, downregulated in hepatocel- lular carcinoma and correlated with prognosis, suppresses cell proliferation and tumourige- nicity. Br. J. Cancer 103:1215-1220.
23. Wang, S., A.B. Aurora, B.A. Johnson, X. Qi, J. McAnally, J.A. Hill, J.A. Richardson, R. Bassel-Duby, and E.N. Olson. 2008. The endothelial-specific microRNA miR-126 governs vascular integrity and angiogenesis. Dev. Cell 15:261-271.
24. Yu, L., N.W. Todd, L. Xing, Y. Xie, H. Zhang, Z. Liu, H. Fang, J. Zhang, et al. 2010. Early detection of lung adenocarcinoma in sputum by a panel of microRNA markers. Int. J. Cancer 127:2870-2878.
25. Xing, L., N.W. Todd, L. Yu, H. Fang, and F. Jiang. 2010. Early detection of squamous cell lung cancer in sputum by a panel of microRNA markers. Mod. Pathol. 23:1157-1164.
26. Beatty, M., J. Guduric-Fuchs, E. Brown, S. Bridgett, U. Chakravarthy, R.E. Hogg, and D.A. Simpson. 2014. Small RNAs from plants, bacteria and fungi within the order Hypoc- reales are ubiquitous in human plasma. BMC Genomics 15:933.
27. Dhahbi, J.M., S.R. Spindler, H. Atamna, D. Boffelli, P. Mote, and D.I.K. Martin. 2013. 5´-YRNA fragments derived by processing of transcripts from specific YRNA genes and pseudogenes are abundant in human serum and plasma. Physiol. Genomics 45:990-998.
28. Dhahbi, J.M., S.R. Spindler, H. Atamna, D. Boffelli, and D.I. Martin. 2014. Deep Sequencing of Serum Small RNAs Identifies Patterns of 5´ tRNA Half and YRNA Fragment Expression Associated with Breast Cancer. Biomark Cancer. 6:37-47.
29. Hilscher, C., W. Vahrson, and D.P. Dittmer. 2005. Faster quantitative real-time PCR protocols may lose sensitivity and show increased variability. Nucleic Acids Res. 33:e182.
30. Kermekchiev, M.B., L.I. Kirilova, E.E. Vail, and W.M. Barnes. 2009. Mutants of Taq DNA polymerase resistant to PCR inhibitors allow DNA amplification from whole blood and crude soil samples. Nucleic Acids Res. 37:e40.
31. Salm, E., Y. Zhong, B. Reddy, C. Duarte- Guevara, V. Swaminathan, Y.-S. Liu, and R. Bashir. 2014. Electrical detection of nucleic acid amplification using an on-chip quasi- reference electrode and a PVC REFET. Anal. Chem. 86:6968-6975.
32. Tijsen, A.J., E.E. Creemers, P.D. Moerland, L.J. de Windt, A.C. van der Wal, W.E. Kok, and Y.M. Pinto. 2010. MiR423-5p as a circu- lating biomarker for heart failure. Circ. Res. 106:1035-1039.
Received 23 January 2015; accepted 06 March 2015.
Address correspondence to David Arthur Simpson, Centre for Experimental Medicine, Queen’s Univer- sity Belfast, Institute of Clinical Science, Royal Victo- ria Hospital, Belfast, Northern Ireland, United King- dom. E-mail:
David.Simpson@
qub.ac.uk
To purchase reprints of this article, contact:
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