Fukushima Ryo, Ogura Yuina, Hosokawa Chikako, Watanabe Noriko, Ishikawa Fumihiro, Shibanuma Motoko, Kato Masaru
Devision of Bioanalytical Chemistry, Department of Pharmaceutical Sciences, School of Pharmacy, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo, 142-8555, Japan.
Center for Biotechnology, Showa University, Tokyo, Japan.
Anal Sci. 2025 Feb;41(2):137-143. doi: 10.1007/s44211-024-00687-4. Epub 2024 Dec 9.
Cell-free DNA (cfDNA) is a valuable biomarker for the early detection of genetic diseases and for evaluating treatment efficacy. We developed a rapid and cost-effective purification method for urinary cfDNA using a commercially available DNA purification kit. This method enables the rapid purification (< 20 min) of DNA suitable for use in the polymerase chain reaction (PCR) using only a centrifuge and a heater. Additionally, we discovered that short-chain DNA could be efficiently purified by incorporating a concentration step using cationic particles. Quantitative PCR (qPCR) analysis of the purified DNA demonstrated that use of the developed method effectively decreased the DNA detection limit. Overall, this method enables the rapid and inexpensive purification of DNA, and it is suitable for combination with recent advanced DNA analysis technologies such as qPCR, next-generation sequencing, and mass spectrometry. It is therefore expected to contribute to the early detection of cancer and have a major impact on the medical field.
游离DNA（cfDNA）是用于遗传疾病早期检测和评估治疗效果的一种有价值的生物标志物。我们使用市售DNA纯化试剂盒开发了一种快速且经济高效的尿液cfDNA纯化方法。该方法仅使用离心机和加热器就能快速纯化（<20分钟）适用于聚合酶链反应（PCR）的DNA。此外，我们发现通过使用阳离子颗粒进行浓缩步骤可以有效地纯化短链DNA。对纯化后DNA的定量PCR（qPCR）分析表明，所开发方法的使用有效地降低了DNA检测限。总体而言，该方法能够快速且廉价地纯化DNA，并且适用于与qPCR、下一代测序和质谱等最新先进DNA分析技术相结合。因此，预计它将有助于癌症的早期检测，并对医学领域产生重大影响。
