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A biocomposite-based rapid sampling assay for circulating cell-free DNA in liquid biopsy samples from human cancers.

Scientific reports (2020-09-12)
Bonhan Koo, Eunsung Jun, Huifang Liu, Eo Jin Kim, Yun-Yong Park, Seok-Byung Lim, Song Cheol Kim, Yong Shin
RESUMEN

Cell-free nucleic acids (cfNAs) in liquid biopsy samples are emerging as important biomarkers for cancer diagnosis and monitoring, and for predicting treatment outcomes. Many cfNA isolation methods have been developed recently. However, most of these techniques are time-consuming, complex, require large equipment, and yield low-purity cfNAs because the genetic background of normal cells is amplified during cell lysis, which limits their clinical application. Here, we report a rapid and simple cfNA sampling platform that can overcome the limitations of conventional methods. We synthesised a biocomposite by combining amine-modified diatomaceous earth (DE) and cucurbituril (CB). The biocomposite platform showed high capture efficiency (86.78-90.26%) with genomic DNA and amplified DNA products (777, 525 and 150 bp). The biocomposite platform allowed the isolation of high purity and quantity cfDNAs from the plasma of 13 cancer patients (three colorectal cancer and ten pancreatic cancer samples) without requiring a lysis step or special equipment. The biocomposite platform may be useful to isolate cfNAs for the diagnosis and treatment of cancers in clinical applications.

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Sigma-Aldrich
Diatomaceous earth, powder, suitable for most filtrations
Sigma-Aldrich
3-Aminopropyl(diethoxy)methylsilane, 97%
Sigma-Aldrich
Cucurbit[6]uril hydrate, contains acid of crystalization