Identification of reference microRNAs and suitability of archived hemopoietic samples for robust microRNA expression profiling

Virginie F. Viprey, Maria V. Corrias, Susan A. Burchill

Research output: Contribution to journalArticle

Abstract

In many cancers, including neuroblastoma, microRNA (miRNA) expression profiling of peripheral blood (PB) and bone marrow (BM) may increase understanding of the metastatic process and lead to the identification of clinically informative biomarkers. The quality of miRNAs in PB and BM samples archived in PAXgene™ blood RNA tubes from large-scale clinical studies and the identity of reference miRNAs for standard reporting of data are to date unknown. In this study, we evaluated the reliability of expression profiling of 377 miRNAs using quantitative polymerase chain reaction (qPCR) in PB and BM samples (n = 90) stored at -80°C for up to 5 years in PAXgene™ blood RNA tubes. There was no correlation with storage time and variation of expression for any single miRNA (r <0.50). The profile of miRNAs isolated as small RNAs or co-isolated with small/large RNAs was highly correlated (r = 0.96). The mean expression of all miRNAs and the geNorm program identified miR-26a, miR-28-5p, and miR-24 as the most stable reference miRNAs. This study describes detailed methodologies for reliable miRNA isolation and profiling of PB and BM, including reference miRNAs for qPCR normalization, and demonstrates the suitability of clinical samples archived at -80°C into PAXgene™ blood RNA tubes for miRNA expression studies.

Original languageEnglish
Pages (from-to)566-572
Number of pages7
JournalAnalytical Biochemistry
Volume421
Issue number2
DOIs
Publication statusPublished - Feb 15 2012

Fingerprint

MicroRNAs
Blood
RNA
Bone
Bone Marrow
Polymerase chain reaction
Polymerase Chain Reaction
Biomarkers
Neuroblastoma
Research Design

Keywords

  • Blood
  • Bone marrow
  • Long-term storage
  • MicroRNAs
  • Neuroblastoma
  • PAXgene™ blood RNA tube
  • Reference microRNAs
  • RT-qPCR

ASJC Scopus subject areas

  • Biochemistry
  • Biophysics
  • Molecular Biology
  • Cell Biology

Cite this

Identification of reference microRNAs and suitability of archived hemopoietic samples for robust microRNA expression profiling. / Viprey, Virginie F.; Corrias, Maria V.; Burchill, Susan A.

In: Analytical Biochemistry, Vol. 421, No. 2, 15.02.2012, p. 566-572.

Research output: Contribution to journalArticle

@article{74b9fc9c8f314dd4bff66d4089a5d009,
title = "Identification of reference microRNAs and suitability of archived hemopoietic samples for robust microRNA expression profiling",
abstract = "In many cancers, including neuroblastoma, microRNA (miRNA) expression profiling of peripheral blood (PB) and bone marrow (BM) may increase understanding of the metastatic process and lead to the identification of clinically informative biomarkers. The quality of miRNAs in PB and BM samples archived in PAXgene™ blood RNA tubes from large-scale clinical studies and the identity of reference miRNAs for standard reporting of data are to date unknown. In this study, we evaluated the reliability of expression profiling of 377 miRNAs using quantitative polymerase chain reaction (qPCR) in PB and BM samples (n = 90) stored at -80°C for up to 5 years in PAXgene™ blood RNA tubes. There was no correlation with storage time and variation of expression for any single miRNA (r <0.50). The profile of miRNAs isolated as small RNAs or co-isolated with small/large RNAs was highly correlated (r = 0.96). The mean expression of all miRNAs and the geNorm program identified miR-26a, miR-28-5p, and miR-24 as the most stable reference miRNAs. This study describes detailed methodologies for reliable miRNA isolation and profiling of PB and BM, including reference miRNAs for qPCR normalization, and demonstrates the suitability of clinical samples archived at -80°C into PAXgene™ blood RNA tubes for miRNA expression studies.",
keywords = "Blood, Bone marrow, Long-term storage, MicroRNAs, Neuroblastoma, PAXgene™ blood RNA tube, Reference microRNAs, RT-qPCR",
author = "Viprey, {Virginie F.} and Corrias, {Maria V.} and Burchill, {Susan A.}",
year = "2012",
month = "2",
day = "15",
doi = "10.1016/j.ab.2011.10.022",
language = "English",
volume = "421",
pages = "566--572",
journal = "Analytical Biochemistry",
issn = "0003-2697",
publisher = "Academic Press Inc.",
number = "2",

}

TY - JOUR

T1 - Identification of reference microRNAs and suitability of archived hemopoietic samples for robust microRNA expression profiling

AU - Viprey, Virginie F.

AU - Corrias, Maria V.

AU - Burchill, Susan A.

PY - 2012/2/15

Y1 - 2012/2/15

N2 - In many cancers, including neuroblastoma, microRNA (miRNA) expression profiling of peripheral blood (PB) and bone marrow (BM) may increase understanding of the metastatic process and lead to the identification of clinically informative biomarkers. The quality of miRNAs in PB and BM samples archived in PAXgene™ blood RNA tubes from large-scale clinical studies and the identity of reference miRNAs for standard reporting of data are to date unknown. In this study, we evaluated the reliability of expression profiling of 377 miRNAs using quantitative polymerase chain reaction (qPCR) in PB and BM samples (n = 90) stored at -80°C for up to 5 years in PAXgene™ blood RNA tubes. There was no correlation with storage time and variation of expression for any single miRNA (r <0.50). The profile of miRNAs isolated as small RNAs or co-isolated with small/large RNAs was highly correlated (r = 0.96). The mean expression of all miRNAs and the geNorm program identified miR-26a, miR-28-5p, and miR-24 as the most stable reference miRNAs. This study describes detailed methodologies for reliable miRNA isolation and profiling of PB and BM, including reference miRNAs for qPCR normalization, and demonstrates the suitability of clinical samples archived at -80°C into PAXgene™ blood RNA tubes for miRNA expression studies.

AB - In many cancers, including neuroblastoma, microRNA (miRNA) expression profiling of peripheral blood (PB) and bone marrow (BM) may increase understanding of the metastatic process and lead to the identification of clinically informative biomarkers. The quality of miRNAs in PB and BM samples archived in PAXgene™ blood RNA tubes from large-scale clinical studies and the identity of reference miRNAs for standard reporting of data are to date unknown. In this study, we evaluated the reliability of expression profiling of 377 miRNAs using quantitative polymerase chain reaction (qPCR) in PB and BM samples (n = 90) stored at -80°C for up to 5 years in PAXgene™ blood RNA tubes. There was no correlation with storage time and variation of expression for any single miRNA (r <0.50). The profile of miRNAs isolated as small RNAs or co-isolated with small/large RNAs was highly correlated (r = 0.96). The mean expression of all miRNAs and the geNorm program identified miR-26a, miR-28-5p, and miR-24 as the most stable reference miRNAs. This study describes detailed methodologies for reliable miRNA isolation and profiling of PB and BM, including reference miRNAs for qPCR normalization, and demonstrates the suitability of clinical samples archived at -80°C into PAXgene™ blood RNA tubes for miRNA expression studies.

KW - Blood

KW - Bone marrow

KW - Long-term storage

KW - MicroRNAs

KW - Neuroblastoma

KW - PAXgene™ blood RNA tube

KW - Reference microRNAs

KW - RT-qPCR

UR - http://www.scopus.com/inward/record.url?scp=84859701762&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84859701762&partnerID=8YFLogxK

U2 - 10.1016/j.ab.2011.10.022

DO - 10.1016/j.ab.2011.10.022

M3 - Article

C2 - 22074795

AN - SCOPUS:84859701762

VL - 421

SP - 566

EP - 572

JO - Analytical Biochemistry

JF - Analytical Biochemistry

SN - 0003-2697

IS - 2

ER -