Postgraduate Medical Institute, Faculty of Medical Science, Anglia Ruskin University, Chelmsford, Essex, UK.
Institute of Population Health, Faculty of Medical and Human Sciences, University of Manchester, Manchester, UK.
Eur J Clin Invest. 2017 Oct;47(10):756-774. doi: 10.1111/eci.12801. Epub 2017 Sep 2.
Poorly executed and inadequately reported molecular measurement methods are amongst the causes underlying the lack of reproducibility of much biomedical research. Although several high impact factor journals have acknowledged their past failure to scrutinise adequately the technical soundness of manuscripts, there is a perplexing reluctance to implement basic corrective measures. The reverse transcription real-time quantitative PCR (RT-qPCR) is probably the most straightforward measurement technique available for RNA quantification and is widely used in research, diagnostic, forensic and biotechnology applications. Despite the impact of the minimum information for the publication of quantitative PCR experiments (MIQE) guidelines, which aim to improve the robustness and the transparency of reporting of RT-qPCR data, we demonstrate that elementary protocol errors, inappropriate data analysis and inadequate reporting continue to be rife and conclude that the majority of published RT-qPCR data are likely to represent technical noise.
执行不当和报告不充分的分子测量方法是许多生物医学研究缺乏可重复性的原因之一。尽管一些高影响因子期刊已经承认过去未能充分审查手稿的技术合理性，但令人费解的是，他们不愿意采取基本的纠正措施。逆转录实时定量 PCR（RT-qPCR）可能是最直接的用于 RNA 定量的测量技术，广泛应用于研究、诊断、法医和生物技术应用中。尽管最小信息发布定量 PCR 实验指南（MIQE）旨在提高 RT-qPCR 数据报告的稳健性和透明度，但我们仍证明基本的协议错误、不适当的数据分析和不充分的报告仍然很普遍，并得出结论，大多数已发表的 RT-qPCR 数据可能代表技术噪声。
