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用于快速灵敏检测循环肿瘤DNA的简单低成本血浆游离核酸采样方法

Simple and Low-Cost Sampling of Cell-Free Nucleic Acids from Blood Plasma for Rapid and Sensitive Detection of Circulating Tumor DNA.

作者信息

Jin Choong Eun, Koo Bonhan, Lee Tae Yoon, Han Kyudong, Lim Seok Byung, Park In Ja, Shin Yong

机构信息

Department of Convergence Medicine Asan Medical Center University of Ulsan College of Medicine 88 Olympicro-43gil, Songpa-gu ,05505 Seoul Republic of Korea.

Biomedical Engineering Research Center Asan Institute of Life Sciences Asan Medical Center 88 Olympicro-43gil, Songpa-gu ,05505 Seoul Republic of Korea.

出版信息

Adv Sci (Weinh). 2018 Jul 30;5(10):1800614. doi: 10.1002/advs.201800614. eCollection 2018 Oct.

Abstract

Cell-free nucleic acids (cfNAs) are emerging diagnostic biomarkers for monitoring the treatment and recurrence of cancers. In particular, the biological role and clinical usefulness of cfNAs obtained from the plasma of patients with various cancers are popular and still intensely explored, yet most studies are limited by technical problems during cfNA isolation. A dimethyl dithiobispropionimidate (DTBP)-based microchannel platform that enables spontaneous cfNA capture in 15 min with minimal cellular background and no requirements for use of bulky instruments is reported first. This platform identified and hot-spot mutations following cfDNA isolation from the blood plasma and tissues obtained from 30 colorectal cancer patients. The correlation of mutations between the primary tissues and plasma from the patients was high using this platform with whole genome sequencing compared to the spin-column method. This platform can also be combined with various detection approaches (biooptical sensor, Sanger sequencing, and polymerase chain reaction (PCR)) for rapid, simple, low-cost, and sensitive circulating tumor DNA detection in blood plasma. The efficiency and versatility of this platform in isolating cfNAs from liquid biopsies has applications in cancer treatment and precision medicine.

摘要

游离核酸(cfNAs)正在成为用于监测癌症治疗和复发的诊断生物标志物。特别是,从各种癌症患者血浆中获得的cfNAs的生物学作用和临床实用性受到广泛关注且仍在深入探索,但大多数研究受到cfNA分离过程中技术问题的限制。首次报道了一种基于二甲基二硫代双丙基亚胺(DTBP)的微通道平台,该平台能够在15分钟内实现自发cfNA捕获,细胞背景极少,且无需使用大型仪器。该平台从30例结直肠癌患者的血浆和组织中分离cfDNA后,鉴定出了热点突变。与柱式离心法相比,使用该平台进行全基因组测序时,患者原发组织和血浆之间的突变相关性很高。该平台还可与各种检测方法(生物光学传感器、桑格测序和聚合酶链反应(PCR))相结合,用于在血浆中快速、简单、低成本且灵敏地检测循环肿瘤DNA。该平台从液体活检中分离cfNAs的效率和多功能性在癌症治疗和精准医学中具有应用价值。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/7ac1/6193143/9a93698eb995/ADVS-5-1800614-g001.jpg

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