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2014 ; 15
(1
): 395
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Comparison of different extraction techniques to profile microRNAs from human
sera and peripheral blood mononuclear cells
#MMPMID24885883
Monleau M
; Bonnel S
; Gostan T
; Blanchard D
; Courgnaud V
; Lecellier CH
BMC Genomics
2014[May]; 15
(1
): 395
PMID24885883
show ga
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.