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基因表达测量的可重复性和可靠性分析

Reproducibility and reliability assays of the gene expression-measurements.

作者信息

Darbani Behrooz, Stewart Charles Neal

机构信息

Department of Plant and Environmental Sciences, University of Copenhagen, Thorvaldsensvej 40, 1871 Frederiksberg, Denmark ; Department of Molecular Biology and Genetics, Research Centre Flakkebjerg, Aarhus University, Forsøgsvej 1, 4200 Slagelse, Denmark.

Department of Plant Sciences, University of Tennessee-Knoxville, 252 Ellington Plant Sciences, 37996-4561 Knoxville, TN USA.

出版信息

J Biol Res (Thessalon). 2014 May 13;21(1):3. doi: 10.1186/2241-5793-21-3. eCollection 2014 Dec.

Abstract

BACKGROUND

Reliability and reproducibility are key metrics for gene expression assays. This report assesses the utility of the correlation coefficient in the analysis of reproducibility and reliability of gene expression data.

RESULTS

The correlation coefficient alone is not sufficient to assess equality among sample replicates but when coupled with slope and scatter plots expression data equality can be better assessed. Narrow-intervals of scatter plots should be shown as a tool to inspect the actual level of noise within the data. Here we propose a method to examine expression data reproducibility, which is based on the ratios of both the means and the standard deviations for the inter-treatment expression ratios of genes. In addition, we introduce a fold-change threshold with an inter-replicate occurrence likelihood lower than 5% to perform analysis even when reproducibility is not acceptable. There is no possibility to find a perfect correlation between transcript and protein levels even when there is not any post-transcriptional regulatory mechanism. We therefore propose an adjustment for protein abundance with that of transcript abundance based on open reading frame length.

CONCLUSIONS

Here, we introduce a very efficient reproducibility approach. Our method detects very small changes in large datasets which was not possible through regular correlation analysis. We also introduce a correction on protein quantities which allows us to examine the post-transcriptional regulatory effects with a higher accuracy.

摘要

背景

可靠性和可重复性是基因表达分析的关键指标。本报告评估了相关系数在基因表达数据可重复性和可靠性分析中的效用。

结果

仅相关系数不足以评估样本重复之间的一致性,但与斜率和散点图结合时,可更好地评估表达数据的一致性。应将散点图的窄区间作为检查数据中实际噪声水平的工具展示出来。在此,我们提出一种基于基因处理间表达比率的均值和标准差比率来检查表达数据可重复性的方法。此外,我们引入了一个重复间出现可能性低于5%的倍数变化阈值,以便即使在可重复性不可接受时也能进行分析。即使不存在任何转录后调控机制,转录本水平与蛋白质水平之间也不可能找到完美的相关性。因此,我们基于开放阅读框长度提出了一种根据转录本丰度调整蛋白质丰度的方法。

结论

在此,我们引入了一种非常有效的可重复性方法。我们的方法能检测大型数据集中非常小的变化,这是常规相关分析无法做到的。我们还对蛋白质量进行了校正,从而使我们能够更准确地检查转录后调控效应。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3f10/4376515/f38c65164dcc/40709_2013_3_Fig1_HTML.jpg

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