title: External oligonucleotide standards enable cross laboratory comparison and exchange of real-time quantitative PCR data.
abstract: 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.
authors.affiliation: Center for Medical Genetics, Ghent University Hospital, Belgium.
authors.forename: Joëlle
authors.initials: J
authors.lastname: Vermeulen
authors.forename: Filip
authors.initials: F
authors.lastname: Pattyn
authors.forename: Katleen
authors.initials: K
authors.lastname: De Preter
authors.forename: Liesbeth
authors.initials: L
authors.lastname: Vercruysse
authors.forename: Stefaan
authors.initials: S
authors.lastname: Derveaux
authors.forename: Pieter
authors.initials: P
authors.lastname: Mestdagh
authors.forename: Steve
authors.initials: S
authors.lastname: Lefever
authors.forename: Jan
authors.initials: J
authors.lastname: Hellemans
authors.forename: Frank
authors.initials: F
authors.lastname: Speleman
authors.forename: Jo
authors.initials: J
authors.lastname: Vandesompele
mesh_terms: D002138:Calibration; D017931:DNA Primers / Q000592:standards*; D015870:Gene Expression; D006801:Humans; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000592:standards*
chemical_list: D017931:DNA Primers