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A DNA-modified hydrogel for simultaneous purification, concentration and detection of targeted cfDNA in human serum

机译：DNA修饰的水凝胶，可同时纯化，浓缩和检测人血清中的靶向cfDNA

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Cell-free DNA (cfDNA) in blood, which stems from the fetus of pregnant women and tumor in cancer patients, has gained attention in molecular diagnosis. However, cfDNA is less stable, and its amount in the serum is extremely low; these are critical barriers for the utilization of this resource. In this study, a DNA-modified polyacrylamide hydrogel (DNA-Gel) was prepared, and a specialized device was designed to simultaneously catch, purify, concentrate, and detect targeted cfDNA by electrophoresis. We demonstrated that 20–1000 bp ssDNA and dsDNA could be caught and released by the DNA-Gel-based device with high specificity and sensitivity. Upon increasing the number of cycles and electrophoresis time, higher DNA purity and density were achieved, and the separation of serum proteins, untargeted cfDNA, and other charged molecules was promoted. As low as 10 pg μL ~(?1) of DNA could be detected using the DNA-Gel after four cycles of concentration. We also detected 1 fg μL ~(?1) of DNA in the serum with 16 cycles of concentration, followed by 25 PCR cycles. We also designed a device to obtain DNA from the DNA-Gel. We found that the DNA loss rate was around 50%, and A260/A280 was close to 1.7. Thus, we have designed a cost-effective and highly economical device to purify DNA at low concentrations with high specificity and selectivity.
机译：血液中的无细胞DNA（cfDNA）源于孕妇的胎儿和癌症患者的肿瘤，在分子诊断中已引起关注。但是，cfDNA的稳定性较差，其在血清中的含量极低。这些是利用该资源的关键障碍。在这项研究中，制备了DNA修饰的聚丙烯酰胺水凝胶（DNA-Gel），并设计了一种专用设备，可通过电泳同时捕获，纯化，浓缩和检测目标cfDNA。我们证明了基于DNA-Gel的设备可以捕获和释放20–1000 bp的ssDNA和dsDNA，并且具有很高的特异性和敏感性。随着循环次数和电泳时间的增加，获得了更高的DNA纯度和密度，并促进了血清蛋白，未靶向cfDNA和其他带电分子的分离。经过四个浓缩周期后，使用DNA-Gel可以检测到低至10 pgμL〜（？1）的DNA。我们还检测了血清中1 fgμL〜（？1）的DNA，浓度为16个循环，然后进行25个PCR循环。我们还设计了一种从DNA-凝胶获得DNA的设备。我们发现DNA丢失率约为50％，而A260 / A280接近1.7。因此，我们设计了一种经济高效且经济的设备，以高特异性和高选择性纯化低浓度的DNA。

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