Optimization of a Protocol for Isolating Cell-free DNA From Cerebrospinal Fluidopen access
- Authors
- Song, Ho Hyun; Park, Hyeran; Cho, Doohwan; Bang, Hae In; Oh, Hyuk-Jin; Kim, Jieun
- Issue Date
- May-2024
- Publisher
- KOREAN SOC LABORATORY MEDICINE
- Keywords
- Cell-free nucleic acids; Cerebrospinal fluid; Circulating tumor DNA; Droplet digi- tal PCR; Vacuum
- Citation
- ANNALS OF LABORATORY MEDICINE, v.44, no.3, pp 294 - 298
- Pages
- 5
- Journal Title
- ANNALS OF LABORATORY MEDICINE
- Volume
- 44
- Number
- 3
- Start Page
- 294
- End Page
- 298
- URI
- https://scholarworks.bwise.kr/sch/handle/2021.sw.sch/26084
- DOI
- 10.3343/alm.2023.0267
- ISSN
- 2234-3806
2234-3814
- Abstract
- A standardized protocol for the isolation of cell-free DNA (cfDNA) from cerebrospinal fluid (CSF) is lacking. Therefore, we established a cfDNA isolation protocol optimized for clinical CSF specimens, integrating acceptable modifications and using artificial CSF generated from remnant CSF spiked with reference cell-free tumor DNA (ctDNA). We compared the isolation yields of in vitro diagnostic (IVD)-certified column-based (CB) and magnetic beadbased (MB) isolation. Furthermore, we modified both methods, including pre- and postelution steps. To confirm ctDNA integrity and quantify the variant allele frequency after isolation, we performed droplet digital PCR (ddPCR) targeting IDH1 R132C in the reference ctDNA. MB isolation had a higher yield than CB isolation (P < 0.0001), and post-isolation vacuum increased the final concentration in both methods, with little effect on cfDNA integrity. Our study provides a protocol to maximize CSF-ctDNA concentrations in IVD testing and future studies.
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Collections - College of Medicine > Department of Anesthesiology > 1. Journal Articles
- College of Medicine > Department of Neurosurgery > 1. Journal Articles
- College of Medicine > Department of Neurosurgery > 1. Journal Articles
- College of Medicine > Department of Clinical Pathology > 1. Journal Articles
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