Detection of ctDNA from Dried Blood Spots after DNA Size Selection.
BACKGROUND: Recent advances in the study and clinical applications of circulating tumor DNA (ctDNA) are limited by practical considerations of sample collection. Whole-genome sequencing (WGS) is increasingly used for analysis of ctDNA, identifying copy-number alterations and fragmentation patterns....
| Publicado en: | Clinical Chemistry Vol. 66; no. 5; pp. 697 - 706 |
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| Autores principales: | , , , , , , , , , , , |
| Formato: | Journal Article |
| Publicado: |
Oxford University Press / USA
May2020
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| Acceso en línea: | Ver este registro en EBSCOhost |
| fields | @attributes: recordID: 1 pdfLink: plink: https://search.ebscohost.com/login.aspx?direct=true&db=ccm&AN=147057604&site=ehost-live header: @attributes: shortDbName: ccm uiTerm: 147057604 longDbName: CINAHL Complete uiTag: AN controlInfo: bkinfo: dissinfo: jinfo: jid: 00099147 10CS jtl: Clinical Chemistry issn: 00099147 maglogo: N pubinfo: dt: May2020 vid: 66 iid: 5 pid: 622 pub: Oxford University Press / USA artinfo: ui: 147057604 10.1093/clinchem/hvaa050 147057604 ppf: 697 ppct: 9 formats: fmt: @attributes: type: P tig: atl: Detection of ctDNA from Dried Blood Spots after DNA Size Selection. aug: au: Heider, Katrin Wan, Jonathan C. M. Hall, James Belic, Jelena Boyle, Samantha Hudecova, Irena Gale, Davina Cooper, Wendy N. Corrie, Pippa G. Brenton, James D. Smith, Christopher G. Rosenfeld, Nitzan affil: Cancer Research UK Cambridge Institute, University of Cambridge, Cambridge, UK sug: ab: BACKGROUND: Recent advances in the study and clinical applications of circulating tumor DNA (ctDNA) are limited by practical considerations of sample collection. Whole-genome sequencing (WGS) is increasingly used for analysis of ctDNA, identifying copy-number alterations and fragmentation patterns. We hypothesized that low-depth/shallow WGS (sWGS) data may be generated from minute amounts of cell-free DNA, and that fragment-size selection may remove contaminating genomic DNA from small blood volumes. Dried blood spots have practical advantages for sample collection, may facilitate serial sampling, and could support novel study designs in humans and animal models. METHODS: We developed a protocol for the isolation and analysis of cell-free DNA from dried blood spots using filter paper cards and bead-based size selection. DNA extracted and size-selected from dried spots was analyzed using sWGS and polymerase chain reaction (PCR). RESULTS: Analyzing a 50 lL dried blood spot from frozen whole blood of a patient with melanoma, we identified ctDNA based on the presence of tumorspecific somatic copy-number alterations, and found a fragment-size profile similar to that observed in plasma DNA. We found alterations in different chromosomes in blood spots from 2 patients with high-grade serous ovarian carcinoma. Extending this approach to serial dried blood spots from mouse xenograft models, we detect tumor-derived cell-free DNA and identified ctDNA from the originally grafted ascites. CONCLUSION: Our data suggest that ctDNA can be detected and monitored in dried blood spots from archived and fresh blood samples, enabling new approaches for sample collection and novel study/trial designs for both patients and in vivo models. pubtype: Academic Journal doctype: Journal Article ougenre: Article language: English refInfo: holdings: @attributes: islocal: N |
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