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从固定组织中提取 DNA 用于下一代测序分析的前分析阶段的标准化。

Standardization of the preanalytical phase of DNA extraction from fixed tissue for next-generation sequencing analyses.

机构信息

IBBL (Integrated BioBank of Luxembourg), 1 rue Louis Rech, L-3555 Dudelange, Luxembourg.

LIH (Luxembourg Institute for Health), 1 A-B Rue Thomas Edison, 1445 Strassen, Luxembourg.

出版信息

N Biotechnol. 2020 Jan 25;54:52-61. doi: 10.1016/j.nbt.2019.07.005. Epub 2019 Aug 6.

Abstract

Next-generation sequencing (NGS) analyses on DNA derived from archived Formalin-Fixed Paraffin-Embedded (FFPE) clinical material can provide a powerful tool in oncology research and clinical diagnostics. Although several studies have established that NGS can be performed using DNA from FFPE tissue, the accuracy and reproducibility of such analyses, as well as their robustness to the biomolecular quality of the samples used, remains a matter of debate. Excellent reviews have recently been published, providing evidence-based best practices for FFPE DNA extraction. Alternative fixatives exist, although their implementation in clinical practice is difficult. In this article, we present (i) a review of fixed tissue DNA preanalytics with a special focus on DNA extraction and fixed tissue sample qualification and (ii) results from comparisons between different methods of DNA extraction from tissue samples that have been fixed or stabilized by different methods, in terms of NGS metrics and different DNA quality metrics.

摘要

下一代测序(NGS)分析从存档的福尔马林固定石蜡包埋(FFPE)临床标本中提取的 DNA,可以为肿瘤学研究和临床诊断提供强大的工具。尽管有几项研究已经证实可以使用 FFPE 组织中的 DNA 进行 NGS,但这些分析的准确性和重现性,以及它们对所使用样本的生物分子质量的稳健性,仍然存在争议。最近发表了一些优秀的综述,为 FFPE DNA 提取提供了循证最佳实践。虽然存在其他固定剂,但在临床实践中实施起来很困难。在本文中,我们介绍了(i)固定组织 DNA 前分析的综述,特别关注 DNA 提取和固定组织样本的质量评估,以及(ii)从不同方法固定或稳定的组织样本中提取 DNA 的不同方法之间的比较结果,比较的指标有 NGS 指标和不同的 DNA 质量指标。

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