Circulating Cell-Free DNA Extraction from Liquid Biopsy for Cancer Research.

IF 1.1 4区 医学 Q3 BIOLOGY Folia Biologica Pub Date : 2022-01-01 DOI:10.14712/fb2022068040153
L Pfeiferova, M Safarikova, J Ulrych, Z Krska, V Frankova, T Zima, M Kalousova
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Abstract

As the number of cancer patients globally increases, a need for reliable biomarkers including circulating tumour DNA from liquid biopsy for diagnosis, prognosis and monitoring of the disease is rising. Currently, mainly tissue samples from biopsy are used, but there are certain limitations: firstly, it is an invasive technique, and secondly, in some cases it is almost impossible to obtain an acceptable tissue sample. This could be changed by using circulating cell-free DNA from liquid biopsy, which also gives the possibility of repeated examination. Here, we focus on the options of isolating circulating cell-free DNA from plasma samples using two isolation techniques: precision manual QIAamp Circulating Nucleic Acid Kit and automatic MagNA Pure Compact (MPC) using Nucleic Acid Isolation Kit I. Manual extraction gave significantly better yields of circulating tumour DNA (P < 0.05). This DNA also had less contaminants (organic compounds or proteins). DNA obtained by both tested methods of isolation is suitable for subsequent molecular genetic methods.

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液体活检循环无细胞DNA提取用于癌症研究。
随着全球癌症患者数量的增加,对可靠的生物标志物(包括液体活检中循环肿瘤DNA)的需求正在增加,以用于疾病的诊断、预后和监测。目前,主要使用活检组织样本,但存在一定的局限性:首先,它是一种侵入性技术,其次,在某些情况下,几乎不可能获得可接受的组织样本。这可以通过使用液体活检中循环的无细胞DNA来改变,这也提供了重复检查的可能性。在这里,我们重点研究了两种分离技术从血浆样品中分离循环无细胞DNA的选择:精密手动QIAamp循环核酸试剂盒和使用核酸分离试剂盒i的自动MagNA纯压缩(MPC)分离技术。手工提取的循环肿瘤DNA产量显著提高(P < 0.05)。这种DNA含有较少的污染物(有机化合物或蛋白质)。通过两种分离方法获得的DNA适用于后续的分子遗传学方法。
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来源期刊
Folia Biologica
Folia Biologica 医学-生物学
CiteScore
1.40
自引率
0.00%
发文量
5
审稿时长
3 months
期刊介绍: Journal of Cellular and Molecular Biology publishes articles describing original research aimed at the elucidation of a wide range of questions of biology and medicine at the cellular and molecular levels. Studies on all organisms as well as on human cells and tissues are welcome.
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