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定量实时PCR实验所需的最少信息。

Minimum information necessary for quantitative real-time PCR experiments.

作者信息

Johnson Gemma, Nour Afif Abdel, Nolan Tania, Huggett Jim, Bustin Stephen

机构信息

Blizard Institute of Cellular and Molecular Science, Queen Mary University, London, UK.

出版信息

Methods Mol Biol. 2014;1160:5-17. doi: 10.1007/978-1-4939-0733-5_2.

Abstract

The MIQE (minimum information for the publication of quantitative real-time PCR) guidelines were published in 2009 with the twin aims of providing a blueprint for good real-time quantitative polymerase chain reaction (qPCR) assay design and encouraging the comprehensive reporting of qPCR protocols. It had become increasingly clear that variable pre-assay conditions, poor assay design, and incorrect data analysis were leading to the routine publication of data that were often inconsistent, inaccurate, and wrong. The problem was exacerbated by a lack of transparency of reporting, with the details of technical information inadequate for the purpose of assessing the validity of published qPCR data. This had, and continues to have serious implications for basic research, reducing the potential for translating findings into valuable applications and potentially devastating consequences for clinical practice. Today, the rationale underlying the MIQE guidelines has become widely accepted, with more than 2,200 citations by March 2014 and editorials in Nature and related publications acknowledging the enormity of the problem. However, the problem we now face is rather serious: thousands of publications that report suspect data are populating and corrupting the peer-reviewed scientific literature. It will be some time before the many contradictions apparent in every area of the life sciences are corrected.

摘要

MIQE(实时定量PCR实验发表的最低信息要求)指南于2009年发布,其双重目标是为良好的实时定量聚合酶链反应(qPCR)实验设计提供蓝图,并鼓励全面报告qPCR实验方案。越来越明显的是,实验前条件多变、实验设计不佳以及数据分析错误导致了常规发表的数据往往不一致、不准确且错误。报告缺乏透明度使问题更加严重,技术信息细节不足以评估已发表qPCR数据的有效性。这已经并将继续对基础研究产生严重影响,降低将研究结果转化为有价值应用的可能性,并可能给临床实践带来毁灭性后果。如今,MIQE指南的基本原理已被广泛接受,截至2014年3月已有超过2200次引用,《自然》及相关出版物的社论也承认了该问题的严重性。然而,我们现在面临的问题相当严重:数千篇报告可疑数据的出版物充斥并破坏着同行评议的科学文献。生命科学各个领域中明显存在的诸多矛盾要得到纠正尚需时日。

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