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Critical appraisal of quantitative PCR results in colorectal cancer research: Can we rely on published qPCR results?
journal contribution
posted on 2023-07-26, 13:30 authored by Jeroen R. Dijkstra, Léon C. Van Kempen, Iris D. Nagtegaal, Stephen A. BustinThe use of real-time quantitative polymerase chain reaction (qPCR) in cancer research has become ubiquitous. The relative simplicity of qPCR experiments, which deliver fast and cost-effective results, means that each year an increasing number of papers utilizing this technique are being published. But how reliable are the published results? Since the validity of gene expression data is greatly dependent on appropriate normalisation to compensate for sample-to-sample and run-to-run variation, we have evaluated the adequacy of normalisation procedures in qPCR-based experiments. Consequently, we assessed all colorectal cancer publications that made use of qPCR from 2006 until August 2013 for the number of reference genes used and whether they had been validated. Using even these minimal evaluation criteria, the validity of only three percent (6/179) of the publications can be adequately assessed. We describe common errors, and conclude that the current state of reporting on qPCR in colorectal cancer research is disquieting. Extrapolated to the study of cancer in general, it is clear that the majority of studies using qPCR cannot be reliably assessed and that at best, the results of these studies may or may not be valid and at worst, pervasive incorrect normalisation is resulting in the wholesale publication of incorrect conclusions. This survey demonstrates that the existence of guidelines, such as MIQE, is necessary but not sufficient to address this problem and suggests that the scientific community should examine its responsibility and be aware of the implications of these findings for current and future research.
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Refereed
- Yes
Volume
8Issue number
4Page range
813-818Publication title
Molecular OncologyISSN
1878-0261External DOI
Publisher
WileyLanguage
- other
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Legacy posted date
2014-01-30Legacy Faculty/School/Department
ARCHIVED Faculty of Health, Social Care & Education (until September 2018)Usage metrics
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