Wei Jia, Zhao Zhen, Gao Jiaxue, Wang Yaoqi, Ma Lina, Meng Xianying, Wang Zhenxin
Department of Thyroid Surgery, The First Hospital of Jilin University, Changchun, Jilin 130021, P. R. China.
State Key Laboratory of Electroanalytical Chemistry, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022, P. R. China.
ACS Omega. 2020 Mar 4;5(10):5365-5371. doi: 10.1021/acsomega.9b04397. eCollection 2020 Mar 17.
A facile method has been developed for the rapid and efficient enrichment of DNAs from different media including synthetic single-strand DNAs (ssDNAs) from buffer solutions and cell-free DNAs (cfDNAs) from blood plasma through electric field-driven adsorption and desorption of DNAs by a polyacrylamide/phytic acid/polydopamine (PAAM/PA/PDA) hydrogel. The as-prepared PAAM/PA/PDA hydrogel possesses regular porosity with a large surface area, strong electric field responsiveness/good conductivity, and a rich aromatic structure, which can be used as an ideal adsorbent for DNA enrichment under a positive electric field. The enriched DNAs can be released efficiently when the positive electric field is converted to a negative electric field. The PAAM/PA/PDA hydrogel-based electrochemical method enables the completion of the process of DNA adsorption and release within 5 min and exhibits reasonable enrichment efficiencies and recovery rates of various DNAs. For instance, the high enrichment sensitivity (0.1 pmol L) together with the excellent recovery (>75%) of an ssDNA with 78 nucleotides is obtained. Combined with the PCR amplification technique, the practicability of the as-proposed method is demonstrated by the screening of circulating tumor DNAs (ctDNAs) with a BRAF mutation in cfDNAs from the blood plasma samples of patients with papillary thyroid cancer or thyroid nodule and random patients from a clinical laboratory.
已经开发出一种简便的方法,通过电场驱动的聚丙烯酰胺/植酸/聚多巴胺(PAAM/PA/PDA)水凝胶对DNA的吸附和解吸作用,从不同介质中快速高效地富集DNA,包括缓冲溶液中的合成单链DNA(ssDNA)和血浆中的游离DNA(cfDNA)。所制备的PAAM/PA/PDA水凝胶具有规则的孔隙率、大表面积、强电场响应性/良好的导电性以及丰富的芳香结构,可作为正电场下DNA富集的理想吸附剂。当正电场转换为负电场时,富集的DNA可以高效释放。基于PAAM/PA/PDA水凝胶的电化学方法能够在5分钟内完成DNA吸附和释放过程,并且对各种DNA表现出合理的富集效率和回收率。例如,对于78个核苷酸的ssDNA,获得了高富集灵敏度(0.1 pmol L)以及优异的回收率(>75%)。结合聚合酶链反应(PCR)扩增技术,通过对甲状腺乳头状癌患者或甲状腺结节患者以及临床实验室随机患者血浆样本中的cfDNA进行BRAF突变循环肿瘤DNA(ctDNA)筛查,证明了所提出方法的实用性。