短 DNA 特异性阻断剂 PCR 可有效且简单地富集短长度的无细胞胎儿 DNA,Chinese Journal of Chemistry

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短 DNA 特异性阻断剂 PCR 可有效且简单地富集短长度的无细胞胎儿 DNA
Chinese Journal of Chemistry ( IF 5.5 ) Pub Date : 2021-04-09 , DOI: 10.1002/cjoc.202100187
Yangwei Liao ₁ , Xiaofeng Tang ₁ , Zhihao Ming ₁ , Lida Ren ₁ , Wei Zhang ₁ , Xianjin Xiao _{1,

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Affiliation

DNA长度作为一种先天特征越来越受到人们的关注。一些重要的生物标志物如ctDNA（循环肿瘤DNA）和cffDNA（无细胞胎儿DNA）的长度比无细胞DNA短。研究人员利用长度差异来增加总游离 DNA 中 ctDNA 或 cffDNA 的丰度，以克服由于丰度低而导致的检测困难。他们开发了短链DNA富集技术，弥补了基于DNA序列差异的典型富集技术应用范围的盲点。然而，现有的短链DNA富集技术不能同时具备高效和简单的特点。在此处，我们开发了 SSB-PCR（Short-DNA Specific Blocker PCR）技术，以高效简单的方式富集短链 DNA。通过理论和实验验证，证明该方法可以将短链DNA的丰度提高10倍以上。基于此，我们找到了该方法在cffDNA富集中的应用场景。此外，还建立了一个程序来对 cffDNA 样本的基因型进行可靠的诊断。我们相信这种方法在实现准确、快速和具有成本效益的无创产前检测方面具有巨大潜力。我们已经找到了该方法在 cffDNA 富集中的应用场景。此外，还建立了一个程序来对 cffDNA 样本的基因型进行可靠的诊断。我们相信这种方法在实现准确、快速和具有成本效益的无创产前检测方面具有巨大潜力。我们已经找到了该方法在 cffDNA 富集中的应用场景。此外，还建立了一个程序来对 cffDNA 样本的基因型进行可靠的诊断。我们相信这种方法在实现准确、快速和具有成本效益的无创产前检测方面具有巨大潜力。

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Short-DNA Specific Blocker PCR for Efficient and Simple Enrichment of Cell Free Fetal DNAs with Short Lengths

As an innate characteristic, DNA length has attracted more and more attention. The length of some important biomarkers such as ctDNA (Circulating tumor DNA) and cffDNA (Cell free fetal DNA) is shorter than that of cell-free DNA. Researchers have utilized the difference in length to increase the abundance of ctDNA or cffDNA in total cell-free DNA to overcome the difficulties in detection due to their low abundance. They have developed short-stranded DNA enrichment technology, which complemented the blind spots of the application range of typical enrichment technology based on DNA sequence differences. However, the existing short-stranded DNA enrichment technology cannot possess high efficiency and simplicity simultaneously. Herein, we developed the SSB-PCR (Short-DNA Specific Blocker PCR) technology to enrich the short-stranded DNA in an efficient and simple way. Through theoretical and experimental verification, it was proved that this method could increase the abundance of short-stranded DNA by more than 10 times. Based on this, we have found an application scenario for this method in cffDNA enrichment. Further, a procedure was established to perform a reliable diagnosis of the genotype of cffDNA samples. We believe that this method has great potential to achieve accurate, fast, and cost-effective non-invasive prenatal testing.

更新日期：2021-04-09

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