ArticleBMC bioinformatics2019
Pairwise efficiency: a new mathematical approach to qPCR data analysis increases the precision of the calibration curve assay.
Article in BMC bioinformatics, 2019. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 8 papers.
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Who cites it
8 citing papers in PubMed.
- Analysis of species adulteration in beef sausage using real-time polymerase chain reaction in Makassar, Indonesia.Veterinary world · 2024Article
- Transcriptome-Based Weighted Gene Co-Expression Network Analysis Reveals the Photosynthesis Pathway and Hub Genes Involved in Promoting Tiller Growth under Repeated Drought-Rewatering Cycles in Perennial Ryegrass.Plants (Basel, Switzerland) · 2024Article
- PCR standard curve quantification in an extensive wastewater surveillance program: results from the Dutch SARS-CoV-2 wastewater surveillance.Frontiers in public health · 2023Article
- Comparative Performance of Eight PCR Methods to DetectPathogens (Basel, Switzerland) · 2021Article
- Quantitative PCR of Small Nucleic Acids: Size Matters.ChemistrySelect · 2021Article
- Evaluation of a real-time RT-PCR panel for detection of SARS-CoV-2 in bat guano.Journal of veterinary diagnostic investigation : official publication of the American Association of Veterinary Laboratory Diagnosticians, Inc · 2021Article
- Analysis of the stability of 70 housekeeping genes during iPS reprogramming.Scientific reports · 2020Article
- Critique of the pairwise method for estimating qPCR amplification efficiency: beware of correlated data!BMC bioinformatics · 2020Article
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Authors and funding
4 authors.
Funding
Abstract
backgroundThe real-time quantitative polymerase chain reaction (qPCR) is routinely used for quantification of nucleic acids and is considered the gold standard in the field of relative nucleic acid measurements. The efficiency of the qPCR reaction is one of the most important parameters in data analysis in qPCR experiments. The Minimum Information for publication of Quantitative real-time PCR Experiments (MIQE) guidelines recommends the calibration curve as the method of choice for estimation of qPCR efficiency. The precision of this method has been reported to be between SD = 0.007 (three replicates) and SD = 0.022 (no replicates).
resultsIn this article, we present a novel approach to the analysis of qPCR data which has been obtained by running a dilution series. Unlike previously developed methods, our method, Pairwise Efficiency, involves a new formula that describes pairwise relationships between data points on separate amplification curves and thus enables extensive statistics. The comparison of Pairwise Efficiency with the calibration curve by Monte Carlo simulation shows the two-folds improvement in the precision of estimations of efficiency and gene expression ratios on the same dataset.
conclusionsThe Pairwise Efficiency nearly doubles the precision in qPCR efficiency determinations compared to standard calibration curve method. This paper demonstrates that applications of combinatorial treatment of data provide the improvement of the determination.
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