The Effect of In Vitro Hemolysis on Measurement of Cell-Free DNA
The Effect of In Vitro Hemolysis on Measurement of Cell-Free DNA
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DOI：
10.1373/jalm.2018.027953
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发表时间：
2019-09-01
影响因子：
2
通讯作者：
Yanagihara, Katsunori
中科院分区：
文献类型：
--
作者：
Nishimura, Fumitaka;Uno, Naoki;Yanagihara, Katsunori
背景：抽血过程中的溶血是实验室伪影的常见原因。尽管目前在常规实践中测量循环无细胞肿瘤DNA和胎儿DNA，但尚未研究体外溶血对无细胞DNA（cfDNA）测量的影响。当体外溶血发生时，细胞DNA可以从受损的白色血细胞中释放出来，降低循环肿瘤DNA和胎儿DNA的分数。结果：机械损伤后，血浆中2个参考基因的cfDNA和游离血红蛋白浓度均升高，其中2个参考基因的cfDNA浓度升高最明显，而2个参考基因的cfDNA浓度升高最明显。cfDNA和游离血红蛋白之间观察到明显的相关性。结论：cfDNA浓度在溶血血浆中较高。因此，通过不适当的血液采集技术，在溶血的血浆中，循环肿瘤DNA和胎儿DNA的分数可能被低估。
Background: Hemolysis during blood drawing is a common cause of laboratory artifacts. Although circulating cell-free tumor DNA and fetal DNA are currently measured in routine practice, the effect of in vitro hemolysis on the measurement of cell-free DNA (cfDNA) has not been investigated. When in vitro hemolysis occurs, cellular DNA could be released from damaged white blood cells and reduce the fraction of circulating tumor DNA and fetal DNA.Methods: Blood from healthy individuals was collected and passed through a narrow needle to cause in vitro hemolysis. Plasma was separated before and after mechanical damage, and concentrations of free hemoglobin and cfDNA of 2 reference genes were measured.Results: cfDNA of 2 reference genes and free hemoglobin increased after mechanical damage. A clear correlation between cfDNA and free hemoglobin was observed.Conclusion: cfDNA concentrations are higher in hemolyzed plasma. Therefore, the fraction of circulating tumor DNA and fetal DNA can be underestimated in plasma hemolyzed by inappropriate blood collection techniques.