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Quantification of Cell-Free DNA in Red Blood Cell Units in Different Whole Blood Processing Methods

机译:不同全血处理方法中红细胞单位中无细胞DNA的定量

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摘要

Background. Whole blood donations in Canada are processed by either the red cell filtration (RCF) or whole blood filtration (WBF) methods, where leukoreduction is potentially delayed in WBF. Fresh WBF red blood cells (RBCs) have been associated with increased in-hospital mortality after transfusion. Cell-free DNA (cfDNA) is released by neutrophils prior to leukoreduction, degraded during RBC storage, and is associated with adverse patient outcomes. We explored cfDNA levels in RBCs prepared by RCF and WBF and different storage durations. Methods. Equal numbers of fresh (stored ≤14 days) and older RBCs were sampled. cfDNA was quantified by spectrophotometry and PicoGreen. Separate regression models determined the association with processing method and storage duration and their interaction on cfDNA. Results. cfDNA in 120 RBC units (73 RCF, 47 WBF) were measured. Using PicoGreen, WBF units overall had higher cfDNA than RCF units (p = 0.0010); fresh WBF units had higher cfDNA than fresh RCF units (p = 0.0093). Using spectrophotometry, fresh RBC units overall had higher cfDNA than older units (p = 0.0031); fresh WBF RBCs had higher cfDNA than older RCF RBCs (p = 0.024). Conclusion. Higher cfDNA in fresh WBF was observed compared to older RCF blood. Further study is required for association with patient outcomes.
机译:背景。加拿大的全血捐赠是通过红细胞过滤(RCF)或全血过滤(WBF)方法处理的,在这些方法中,WBF可能会导致白细胞减少。新鲜的WBF红细胞(RBC)与输血后住院死亡率增加有关。无细胞DNA(cfDNA)在白细胞减少之前由嗜中性粒细胞释放,在RBC储存过程中降解,并与患者不良的预后相关。我们探索了由RCF和WBF制备的RBC中的cfDNA水平以及不同的储存时间。方法。抽取了相等数量的新鲜(存储≤14天)和较旧的RBC。通过分光光度法和PicoGreen对cfDNA进行定量。单独的回归模型确定了与加工方法和存储时间的关联以及它们在cfDNA上的相互作用。结果。测量了120个RBC单位(73 RCF,47 WBF)中的cfDNA。使用PicoGreen,WBF单元的总体cfDNA高于RCF单元(p = 0.0010);新鲜的WBF单位比新鲜的RCF单位具有更高的cfDNA(p = 0.0093)。使用分光光度法,新鲜的RBC单元总体上具有比旧单元更高的cfDNA(p = 0.0031);新鲜的WBF RBC比旧的RCF RBC具有更高的cfDNA(p = 0.024)。结论。与较旧的RCF血液相比,新鲜WBF中的cfDNA更高。与患者预后的关联需要进一步的研究。

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