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提高定量聚合酶链反应实验的质量:《MIQE指南》发布15年

Improving the quality of quantitative polymerase chain reaction experiments: 15 years of MIQE.

作者信息

Bustin Stephen A

机构信息

Medical Technology Research Centre, Anglia Ruskin University, Chelmsford, Essex, CM1 1SQ, UK.

出版信息

Mol Aspects Med. 2024 Apr;96:101249. doi: 10.1016/j.mam.2024.101249. Epub 2024 Jan 29.

Abstract

The quantitative polymerase chain reaction (qPCR) is fundamental to molecular biology. It is not just a laboratory technique, qPCR is a bridge between research and clinical practice. Its theoretical foundations guide the design of experiments, while its practical implications extend to diagnostics, treatment, and research advancements in the life sciences, human and veterinary medicine, agriculture, and forensics. However, the accuracy, reliability and reproducibility of qPCR data face challenges arising from various factors associated with experimental design, execution, data analysis and inadequate reporting details. Addressing these concerns, the Minimum Information for the Publication of Quantitative Real-Time PCR Experiments (MIQE) guidelines have emerged as a cohesive framework offering a standardised set of recommendations that describe the essential information required for assessing qPCR experiments. By emphasising the importance of methodological rigour, the MIQE guidelines have made a major contribution to improving the trustworthiness, consistency, and transparency of many published qPCR results. However, major challenges related to awareness, resources, and publication pressures continue to affect their consistent application.

摘要

定量聚合酶链反应(qPCR)是分子生物学的基础。它不仅仅是一种实验室技术,qPCR还是研究与临床实践之间的桥梁。其理论基础指导实验设计,而其实践意义则延伸至诊断、治疗以及生命科学、人类和兽医学、农业及法医学等领域的研究进展。然而,qPCR数据的准确性、可靠性和可重复性面临着来自与实验设计、执行、数据分析以及报告细节不足相关的各种因素所带来的挑战。为解决这些问题,《定量实时PCR实验发表的最低信息要求》(MIQE)指南应运而生,成为一个连贯的框架,提供了一套标准化的建议,描述了评估qPCR实验所需的基本信息。通过强调方法严谨性的重要性,MIQE指南为提高许多已发表的qPCR结果的可信度、一致性和透明度做出了重大贡献。然而,与认知、资源和发表压力相关的重大挑战继续影响着它们的一致应用。

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