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External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data

The quantitative polymerase chain reaction (qPCR) is widely utilized for gene expression analysis. However, the lack of robust strategies for cross laboratory data comparison hinders the ability to collaborate or perform large multicentre studies conducted at different sites. In this study we introd...

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Autores principales: Vermeulen, Joëlle, Pattyn, Filip, De Preter, Katleen, Vercruysse, Liesbeth, Derveaux, Stefaan, Mestdagh, Pieter, Lefever, Steve, Hellemans, Jan, Speleman, Frank, Vandesompele, Jo
Formato: Texto
Lenguaje:English
Publicado: Oxford University Press 2009
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2790878/
https://www.ncbi.nlm.nih.gov/pubmed/19734345
http://dx.doi.org/10.1093/nar/gkp721
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author Vermeulen, Joëlle
Pattyn, Filip
De Preter, Katleen
Vercruysse, Liesbeth
Derveaux, Stefaan
Mestdagh, Pieter
Lefever, Steve
Hellemans, Jan
Speleman, Frank
Vandesompele, Jo
author_facet Vermeulen, Joëlle
Pattyn, Filip
De Preter, Katleen
Vercruysse, Liesbeth
Derveaux, Stefaan
Mestdagh, Pieter
Lefever, Steve
Hellemans, Jan
Speleman, Frank
Vandesompele, Jo
author_sort Vermeulen, Joëlle
collection PubMed
description The quantitative polymerase chain reaction (qPCR) is widely utilized for gene expression analysis. However, the lack of robust strategies for cross laboratory data comparison hinders the ability to collaborate or perform large multicentre studies conducted at different sites. In this study we introduced and validated a workflow that employs universally applicable, quantifiable external oligonucleotide standards to address this question. Using the proposed standards and data-analysis procedure, we obtained a perfect concordance between expression values from eight different genes in 366 patient samples measured on three different qPCR instruments and matching software, reagents, plates and seals, demonstrating the power of this strategy to detect and correct inter-run variation and to enable exchange of data between different laboratories, even when not using the same qPCR platform.
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spelling pubmed-27908782009-12-09 External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data Vermeulen, Joëlle Pattyn, Filip De Preter, Katleen Vercruysse, Liesbeth Derveaux, Stefaan Mestdagh, Pieter Lefever, Steve Hellemans, Jan Speleman, Frank Vandesompele, Jo Nucleic Acids Res Methods Online The quantitative polymerase chain reaction (qPCR) is widely utilized for gene expression analysis. However, the lack of robust strategies for cross laboratory data comparison hinders the ability to collaborate or perform large multicentre studies conducted at different sites. In this study we introduced and validated a workflow that employs universally applicable, quantifiable external oligonucleotide standards to address this question. Using the proposed standards and data-analysis procedure, we obtained a perfect concordance between expression values from eight different genes in 366 patient samples measured on three different qPCR instruments and matching software, reagents, plates and seals, demonstrating the power of this strategy to detect and correct inter-run variation and to enable exchange of data between different laboratories, even when not using the same qPCR platform. Oxford University Press 2009-11 2009-09-04 /pmc/articles/PMC2790878/ /pubmed/19734345 http://dx.doi.org/10.1093/nar/gkp721 Text en © The Author(s) 2009. Published by Oxford University Press. http://creativecommons.org/licenses/by-nc/2.5/uk/ This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/2.5/uk/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Methods Online
Vermeulen, Joëlle
Pattyn, Filip
De Preter, Katleen
Vercruysse, Liesbeth
Derveaux, Stefaan
Mestdagh, Pieter
Lefever, Steve
Hellemans, Jan
Speleman, Frank
Vandesompele, Jo
External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
title External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
title_full External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
title_fullStr External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
title_full_unstemmed External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
title_short External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data
title_sort external oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative pcr data
topic Methods Online
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2790878/
https://www.ncbi.nlm.nih.gov/pubmed/19734345
http://dx.doi.org/10.1093/nar/gkp721
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