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Title:An international comparability study on quantification of mRNA gene expression ratios: CCQM-P103.1
Authors:ID Devonshire, Alison S. (Author)
ID Demšar, Tina (Author)
ID Žel, Jana (Author)
ID Blejec, Andrej (Author)
ID Milavec, Mojca (Author)
Files:.pdf PDF - Presentation file, download (1,78 MB)
MD5: 64D199C43AC0EB7DF78AD7D3CA19BA15
 
URL URL - Source URL, visit https://doi.org/10.1016/j.bdq.2016.05.003
 
Language:English
Typology:1.01 - Original Scientific Article
Organization:Logo NIB - National Institute of Biology
Abstract:Measurement of RNA can be used to study and monitor a range of infectious and non-communicable diseases, with profiling of multiple gene expression mRNA transcripts being increasingly applied to cancer stratification and prognosis. An international comparison study (Consultative Committee for Amount of Substance (CCQM)-P103.1) was performed in order to evaluate the comparability of measurements of RNA copy number ratio for multiple gene targets between two samples. Six exogenous synthetic targets comprising of External RNA Control Consortium (ERCC) standards were measured alongside transcripts for three endogenous gene targets present in the background of human cell line RNA. The study was carried out under the auspices of the Nucleic Acids (formerly Bioanalysis) Working Group of the CCQM. It was coordinated by LGC (United Kingdom) with the support of National Institute of Standards and Technology (USA) and results were submitted from thirteen National Metrology Institutes and Designated Institutes. The majority of laboratories performed RNA measurements using RT-qPCR, with datasets also being submitted by two laboratories based on reverse transcription digital polymerase chain reaction and one laboratory using a next-generation sequencing method. In RT-qPCR analysis, the RNA copy number ratios between the two samples were quantified using either a standard curve or a relative quantification approach. In general, good agreement was observed between the reported results of ERCC RNA copy number ratio measurements. Measurements of the RNA copy number ratios for endogenous genes between the two samples were also consistent between the majority of laboratories. Some differences in the reported values and confidence intervals (‘measurement uncertainties’) were noted which may be attributable to choice of measurement method or quantification approach. This highlights the need for standardised practices for the calculation of fold change ratios and uncertainties in the area of gene expression profiling.
Keywords:RNA copy number ratio, RT-qPCR, gene expression, normalisation, standardisation, molecular diagnostic, transcriptomics, cancer, diagnostics, biomarker identification and validation
Publication status:Published
Publication version:Version of Record
Publication date:01.06.2016
Year of publishing:2016
Number of pages:str. 15-28
Numbering:Vol. 8
PID:20.500.12556/DiRROS-21601 New window
UDC:577.2
ISSN on article:Y507-7052
DOI:10.1016/j.bdq.2016.05.003 New window
COBISS.SI-ID:3906639 New window
Note:Nasl. z nasl. zaslona; Opis vira z dne 9. 6. 2016;
Publication date in DiRROS:04.03.2025
Views:71
Downloads:31
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DEVONSHIRE, Alison S., DEMŠAR, Tina, ŽEL, Jana, BLEJEC, Andrej and MILAVEC, Mojca, 2016, An international comparability study on quantification of mRNA gene expression ratios: CCQM-P103.1. Biomolecular detection and quantification [online]. 2016. Vol. 8, p. 15–28. [Accessed 13 March 2025]. DOI 10.1016/j.bdq.2016.05.003. Retrieved from: https://dirros.openscience.si/IzpisGradiva.php?lang=eng&id=21601
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Record is a part of a journal

Title:Biomolecular detection and quantification
Publisher:Elsevier
COBISS.SI-ID:3411023 New window

Document is financed by a project

Funder:RCUK - Research Council UK
Funding programme:National Measurement System

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License:CC BY-NC-ND 4.0, Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International
Link:http://creativecommons.org/licenses/by-nc-nd/4.0/
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