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Towards clinical applications of blood-born miRNA signatures: The influence of the anticoagulant EDTA on miRNA abundance

ABSTRACT: Circulating microRNAs (miRNAs) from blood are increasingly recognized as biomarker candidates for human diseases. Clinical routine settings frequently include blood sampling in tubes with EDTA as anticoagulant without considering the influence of phlebotomy on the overall miRNA expression pattern. We collected blood samples from six healthy individuals each in an EDTA blood collection tube. Subsequently, the blood was transferred into PAXgeneTM tubes at three different time points, i.e. directly (0 min), 10 min, and 2 h after phlebotomy. As control blood was also directly collected in PAXgeneTM blood RNA tubes that contain a reagent to directly lyse blood cells and stabilize their content. For all six blood donors at the four conditions (24 samples) we analyzed the abundance of 1,205 miRNAs by human Agilent miRNA V16 microarrays.

ORGANISM(S): Homo sapiens

PROVIDER: GSE73401 | GEO | 2015/12/31

SECONDARY ACCESSION(S): PRJNA296893

REPOSITORIES: GEO

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Towards clinical applications of blood-born miRNA signatures: The influence of the anticoagulant EDTA on miRNA abundance

Project description:Circulating microRNAs (miRNAs) from blood are increasingly recognized as biomarker candidates for human diseases. Clinical routine settings frequently include blood sampling in tubes with EDTA as anticoagulant without considering the influence of phlebotomy on the overall miRNA expression pattern. We collected blood samples from six healthy individuals each in an EDTA blood collection tube. Subsequently, the blood was transferred into PAXgeneTM tubes at three different time points, i.e. directly (0 min), 10 min, and 2 h after phlebotomy. As control blood was also directly collected in PAXgeneTM blood RNA tubes that contain a reagent to directly lyse blood cells and stabilize their content. For all six blood donors at the four conditions (24 samples) we analyzed the abundance of 1,205 miRNAs by human Agilent miRNA V16 microarrays. Blood from 6 healthy individuals was collected in one PAXgeneTM blood RNA tube (Becton Dickinson, 2.5 ml blood) and one dipotassium EDTA blood tube (EDTA-KE Monovette, Sarstedt, 9 ml blood) per individual. There was no known disease for any of the blood donors. A fixed volume of 2.5 ml blood from the EDTA tube was transferred at three different time points after blood withdrawal (0 min, 10 min, and 2 h) into fresh PAXgene blood RNA tubes to ensure stabilization of the RNA in the blood samples. All PAXgene blood tubes were stored at room temperature until at least 2 h after the last transfer of EDTA blood into PAXgene tubes, to ensure complete lysis of the blood cells, before they were stored at -20Â°C until RNA isolation.

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