2021
DOI: 10.1155/2021/5585148
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Optimization of Preanalytical Variables for cfDNA Processing and Detection of ctDNA in Archival Plasma Samples

Abstract: DNA released from cells into the peripheral blood is known as cell-free DNA (cfDNA), representing a promising noninvasive source of biomarkers that could be utilized to manage Diffuse Large B-Cell Lymphoma (DLBCL), among other diseases. The procedure for purification and handling of cfDNA is not yet standardized, and various preanalytical variables may affect the yield and analysis of cfDNA, including the purification kits, blood collection tubes, and centrifugation regime. Therefore, we aimed to investigate t… Show more

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Cited by 10 publications
(13 citation statements)
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References 28 publications
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“…To allow standardization, some preanalytical factors involved in the early procedures to separate the liquid part of blood from blood cells require investigation 78 , 79 , 80 and global consensus needs to be sought on the assignment of values and standards across multiple clinical centers and diagnostic laboratories through some standardization alliance. 81 Furthermore, to avoid false-negative detection, a lower limit of detection needs to be determined and a standard curve has to be constructed for quantifying the allelic frequencies of the mutants in the cfDNA.…”
Section: Use Of Cfdna In the Clinical Management Of Hccmentioning
confidence: 99%
“…To allow standardization, some preanalytical factors involved in the early procedures to separate the liquid part of blood from blood cells require investigation 78 , 79 , 80 and global consensus needs to be sought on the assignment of values and standards across multiple clinical centers and diagnostic laboratories through some standardization alliance. 81 Furthermore, to avoid false-negative detection, a lower limit of detection needs to be determined and a standard curve has to be constructed for quantifying the allelic frequencies of the mutants in the cfDNA.…”
Section: Use Of Cfdna In the Clinical Management Of Hccmentioning
confidence: 99%
“…A significantly higher yield of cfDNA was obtained by QIAamp Circulating Nucleic Acid purification kit (Figure 15, p= 0.006570, one way ANOVA test). Blood was collected from six healthy volunteers in EDTA and Streck BCTs, and we observed no significant difference in cfDNA yield purified from plasma separated from Streck and EDTA tubes freshly, after 1 h, and 4 h. However, significantly higher cfDNA yield was observed in plasma samples purified from EDTA BCTs after 24 h of storage at 4°C, which could be due to gDNA contamination [158]. Comparison of centrifugation regimens revealed a slightly higher yield of cfDNA extracted from the plasma of blood samples centrifuged at 2000 x g at RT.…”
Section: Paper IIImentioning
confidence: 60%
“…These samples had matched tumor gDNA that previously was assessed by WES and ddPCR. Patients with tumor biopsies with mutations detected in CD58, TNFRSF14, or EZH2 had detectable ctDNA in the cfDNA assessed with ddPCR and mutation-specific assays (Figure 17) [158]. This demonstrates the usability of archival clinical samples, though they were not processed with fully optimal conditions.…”
Section: Paper IIImentioning
confidence: 99%
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