Email updates

Keep up to date with the latest news and content from BMC Genomics and BioMed Central.

Open Access Methodology article

Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells

Marjorie Monleau1234, Sophie Bonnel1234, Thierry Gostan123, Dominique Blanchard4, Valérie Courgnaud123* and Charles-Henri Lecellier123*

Author Affiliations

1 Institut de Génétique Moléculaire de Montpellier UMR 5535 CNRS, 34293 Montpellier cedex 5, France

2 Université Montpellier 2, Place Eugène Bataillon, 34095 Montpellier cedex 5, France

3 Université Montpellier 1, 5 Bd Henry IV, 34967 Montpellier cedex 2, France

4 Present address: Prestizia/Theradiag group, Cap Alpha, 9 avenue de l’Europe, F-34830 Clapiers, France

For all author emails, please log on.

BMC Genomics 2014, 15:395  doi:10.1186/1471-2164-15-395

Published: 23 May 2014

Abstract

Background

microRNAs (miRNAs) play crucial roles in major biological processes and their deregulations are often associated with human malignancies. As such, they represent appealing candidates as targets of innovative therapies. Another interesting aspect of their biology is that they are present in various biological fluids where, advantageously, they appear to be very stable. A plethora of studies have now reported their potential as biomarkers that can be used in diagnosis, prognosis and/or theranostic issues. However, the application of circulating miRNAs in clinical practices still requires the identification of highly efficient, robust and reproducible methods for their isolation from biological samples.

In that context, we performed an independent cross-comparison of three commercially available RNA extraction kits for miRNAs isolation from human blood samples (Qiagen and Norgen kits as well as the new NucleoSpin miRNAs Plasma kit from Macherey-Nagel). miRNAs were further profiled using the Taqman Low Density Array technology.

Results

We found that, although these 3 kits had equal performances in extracting miRNAs from peripheral blood mononuclear cells, the Macherey-Nagel kit presented several advantages when isolating miRNAs from sera. Besides, our results have indicated that, depending on the quantity of the biological samples used, the extraction procedure directly impacted on the G/C composition of the miRNAs detected.

Conclusion

Overall, our study contributes to the definition of a reliable framework for profiling circulating miRNAs.