A rapid purification method for trace amounts of cell-free DNA in urine

Anal Sci. 2024 Dec 9. doi: 10.1007/s44211-024-00687-4. Online ahead of print.

Abstract

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.

Keywords: Cell-free DNA; Liquid biopsy; Purification method; Urine; qPCR.

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