Selective extraction of low-abundance BRAF V600E mutation from plasma, urine, and sputum using ion-tagged oligonucleotides and magnetic ionic liquids
Selective extraction of low-abundance BRAF V600E mutation from plasma, urine, and sputum using ion-tagged oligonucleotides and magnetic ionic liquids
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DOI：
10.1007/s00216-021-03216-8
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发表时间：
2021-03-01
影响因子：
4.3
通讯作者：
Anderson，Jared L.
中科院分区：
文献类型：
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作者：
Emaus，Miranda N.;Anderson，Jared L.
序列特异性DNA提取有可能改善复杂基质中低丰度突变的检测，使其成为癌症早期循环肿瘤DNA分析的理想选择。离子标记的寡核苷酸（ITO）是通过硫醇烯点击化学用烯丙基咪唑鎓盐修饰的寡核苷酸。基于烯丙基咪唑的标签允许ITO-DNA双链体被疏水磁性离子液体（MIL）选择性地捕获。在这项研究中，ITO-MIL方法的选择性是通过提取低丰度的BRAFV 600 E突变-一种常见的单核苷酸多态性与几种不同的癌症-从稀释的人血浆，人工尿液，稀释的人工痰。定量聚合酶链反应（qPCR）无法区分9%BRAFV 600 E标准品（50 fg·μL− 1 BRAFV 600 E，500 fg·μL− 1野生型BRAF）和100%野生型BRAF（50 fg·μL−1）标准品。然而，在qPCR之前引入ITO-MIL提取允许样品由0.1%BRAFV600E组成（50 fg·μL-1V 600 EBRAF，50，000 fg·μL-1野生型BRAF）以与100%野生型BRAF标准品区分。图形摘要将离子标记的寡核苷酸（ITO）与磁性离子液体（MIL）组合以从稀释的人血浆中提取低丰度BRAFV 600 E突变，人造尿液和稀释的人造痰。在qPCR之前的ITO-MIL提取允许将由0.1%BRAFV600E组成的样品与100%野生型BRAF标准品区分开
Sequence-specific DNA extractions have the potential to improve the detection of low-abundance mutations from complex matrices, making them ideal for circulating tumor DNA analysis during the early stages of cancer. Ion-tagged oligonucleotides (ITOs) are oligonucleotides modified with an allylimidazolium salt via thiolene click chemistry. The allylimidazolium-based tag allows the ITO-DNA duplex to be selectively captured by a hydrophobic magnetic ionic liquid (MIL). In this study, the selectivity of the ITO-MIL method was examined by extracting low abundance of theBRAFV600E mutation—a common single-nucleotide polymorphism associated with several different cancers—from diluted human plasma, artificial urine, and diluted artificial sputum. Quantitative polymerase chain reaction (qPCR) was not able to distinguish a 9%BRAFV600E standard (50 fg·μL−1BRAFV600E, 500 fg·μL−1wild-typeBRAF) from the 100% wild-typeBRAF(50 fg·μL−1) standard. However, introducing the ITO-MIL extraction prior to qPCR allowed for samples consisting of 0.1%BRAFV600E (50 fg·μL−1V600EBRAF, 50,000 fg·μL−1wild-typeBRAF) to be distinguished from the 100% wild-typeBRAFstandard.Graphical abstractIon-tagged oligonucleotides (ITOs) are combined with magnetic ionic liquids (MILs) to extract low-abundanceBRAFV600E mutation from diluted human plasma, artificial urine, and diluted artificial sputum. The ITO-MIL extraction prior to qPCR allowed for samples consisting of 0.1%BRAFV600E to be distinguished from the 100% wild-typeBRAFstandard