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Performance comparison of blood collection tubes as liquid biopsy storage system for minimizing cfDNA contamination from genomic DNA

机译:血液采集管作为液体活检存储系统以最小化基因组DNA对cfDNA污染的性能比较

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Background The ratio of target cfDNA in total plasma is low. The abundant gDNA background resulting from blood cell lysis caused by improper operation has become a major obstacle to accurately measure cfDNA. In this study, we investigated the storage capacity of three blood cell collection tubes (BCTs) in the prevention of genomic DNA contamination caused by white blood cell rupture and evaluated their performance when they were utilized combining with highly sensitive mutation detection technology. Methods Blood samples were drawn from six healthy blood donors and stored in three types of BCTs (BD Ksub2/sub‐EDTA tube, Roche tube, and Streck tube). Plasma samples were isolated at specific time points (day 0, day 3, day 7, and day 14) for content analysis. Results Roche BCT was more capable for preventing cfDNA contamination caused by white blood cell disruption within 14?days, comparing with BD Ksub2/sub‐EDTA tube and Streck tube. Severe white blood cell lysis and gDNA contamination were found in the BD tube. The impacts of Roche and Streck tubes on the quantity and complexity of next‐generation sequencing (NGS) libraries did not differ significantly within 3?days, satisfying most of our daily demands. In addition, the rupture of WBC was not synchronized with hemolysis in BCTs. Conclusion This study showed that capacities of blood collection tubes differed considerably in preservation of blood samples. Therefore, suitable blood collection devices should be selected to minimize gDNA contamination and to standardize blood samples processing for achieving more accurate and reliable clinical analysis of cfDNA.
机译:背景技术靶cfDNA在总血浆中的比例很低。因操作不当而引起的血细胞裂解而产生的丰富gDNA背景已经成为准确测量cfDNA的主要障碍。在这项研究中,我们调查了三个血细胞收集管(BCT)在预防白细胞破裂引起的基因组DNA污染方面的存储能力,并结合高灵敏度突变检测技术评估了它们的性能。方法从六个健康的献血者那里抽取血液样本,并将其存储在三种类型的BCT中(BD K 2 -EDTA管,Roche管和Streck管)。在特定时间点(第0天,第3天,第7天和第14天)分离血浆样品以进行含量分析。结果与BD K 2 -EDTA管和Streck管相比,Roche BCT在14天之内更能防止白细胞破坏引起的cfDNA污染。在BD管中发现严重的白细胞溶解和gDNA污染。罗氏管和Streck管对下一代测序(NGS)文库的数量和复杂性的影响在3天之内变化不大,可以满足我们大多数日常需求。另外,在BCT中,WBC的破裂与溶血不同步。结论这项研究表明,血液采集管在保存血液样本方面存在很大差异。因此,应选择合适的采血设备以最小化gDNA污染并使血液样本处理标准化,以实现cfDNA的更准确和可靠的临床分析。

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