PCR-based methods for the enrichment of minority alleles and mutations.
PCR-based methods for the enrichment of minority alleles and mutations.
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DOI：
10.1373/clinchem.2008.113035
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发表时间：
2009-04
影响因子：
9.3
通讯作者：
Makrigiorgos GM
中科院分区：
文献类型：
--
作者：
Milbury CA;Li J;Makrigiorgos GM
在过量的野生型样本中识别低水平体细胞DNA突变和少数等位基因的能力对于癌症患者早期和治疗后肿瘤状态的表征变得至关重要。在过去的20年里，许多研究都集中在提高pcr技术的选择性上，以增强临床样本中少数（突变）等位基因的检测。在临床和诊断环境中的常规应用要求这些技术是准确的和具有成本效益的，并且需要很少的努力来优化、执行和分析。富集方法通常根据其富集和检测已知或未知突变的能力进行区分。虽然有几种强大的方法可以检测高背景野生型DNA中的已知突变，但很少有技术能够丰富和检测低水平未知突变。一个有希望的发展是COLD-PCR（低变性温度下的共扩增），它可以在任何位置富集含有未知突变的PCR扩增子，这样它们就可以随后测序以确定确切的核苷酸变化。这篇综述总结了用于检测少数DNA突变的技术，重点介绍了有助于富集未知低水平DNA变异的新方法，以便随后对突变进行测序。少数等位基因的富集在临床和诊断应用中是必要的，特别是在与癌症检测有关的应用中，并且持续的技术发展是必要的。
The ability to identify low-level somatic DNA mutations and minority alleles within an excess wild-type sample is becoming essential for characterizing early and posttreatment tumor status in cancer patients. Over the past 2 decades, much research has focused on improving the selectivity of PCR-based technologies for enhancing the detection of minority (mutant) alleles in clinical samples. Routine application in clinical and diagnostic settings requires that these techniques be accurate and cost-effective and require little effort to optimize, perform, and analyze. Enrichment methods typically segregate by their ability to enrich for, and detect, either known or unknown mutations. Although there are several robust approaches for detecting known mutations within a high background of wild-type DNA, there are few techniques capable of enriching and detecting low-level unknown mutations. One promising development is COLD-PCR (coamplification at lower denaturation temperature), which enables enrichment of PCR amplicons containing unknown mutations at any position, such that they can be subsequently sequenced to identify the exact nucleotide change. This review summarizes technologies available for detecting minority DNA mutations, placing an emphasis on newer methods that facilitate the enrichment of unknown low-level DNA variants such that the mutation can subsequently be sequenced. The enrichment of minority alleles is imperative in clinical and diagnostic applications, especially in those related to cancer detection, and continued technology development is warranted.
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影响因子：
2.7
作者：
Kaur, M;Zhang, YZ;Makrigiorgos, GM
通讯作者：
Makrigiorgos, GM
影响因子：
8.8
作者：
通讯作者：
--
影响因子：
14.9
作者：
HALIASSOS, A;CHOMEL, JC;KITZIS, A
通讯作者：
KITZIS, A
影响因子：
14.9
作者：
Amicarelli G;Shehi E;Makrigiorgos GM;Adlerstein D
通讯作者：
Adlerstein D
影响因子：
14.9
作者：
Pincas, H;Pingle, MR;Barany, F
通讯作者：
Barany, F