Comparison of different extraction techniques to profile microRNAs from human sera and peripheral blood mononuclear cells
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
BMC Genomics 2014, 15:395 doi:10.1186/1471-2164-15-395Published: 23 May 2014
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.
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.
Overall, our study contributes to the definition of a reliable framework for profiling circulating miRNAs.