Haemolysis during sample preparation alters microRNA content of plasma.
Haemolysis during sample preparation alters microRNA content of plasma.
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DOI：
10.1371/journal.pone.0024145
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Reid G
中科院分区：
文献类型：
--
作者：
Kirschner MB;Kao SC;Edelman JJ;Armstrong NJ;Vallely MP;van Zandwijk N;Reid G
在一系列体液中已经检测到无细胞microRNA（miRNAs）的存在。特别是血浆/血清中的miRNA含量已被提出作为许多疾病的新型生物标志物的潜在来源。然而，由于分离效率低和缺乏关于最佳参考miRNA的共识，使得来自血浆或血清的miRNA的定量变得困难。溶血对血浆中miRNA的定量和标准化的影响尚未详细研究。我们发现，当在健康志愿者或恶性间皮瘤或冠状动脉疾病患者的血浆样本中测量时，miR-16（一种常用的参考基因）的水平几乎没有变化。包括具有溶血证据的样品导致miR-16水平的变化，因此降低了其作为参考的能力。miR-16和miR-451的水平（两者均以显著水平存在于红细胞中）与溶血程度成比例。对全血、血浆、红细胞和外周血单核细胞中这些miRNA水平的测量显示，红细胞的miRNA含量代表了血浆中测量的miR-16和miR-451水平的主要变异来源。将裂解的红细胞添加到未溶血的血浆中允许测定游离血红蛋白的截止水平，低于该截止水平，个体之间的miR-16和miR-451水平几乎没有变化。总之，血浆miR-16和miR-451的增加由溶血引起。在不存在溶血的情况下，miR-16和miR-451的水平足够恒定以用作标准化物。
The presence of cell-free microRNAs (miRNAs) has been detected in a range of body fluids. The miRNA content of plasma/serum in particular has been proposed as a potential source of novel biomarkers for a number of diseases. Nevertheless, the quantification of miRNAs from plasma or serum is made difficult due to inefficient isolation and lack of consensus regarding the optimal reference miRNA. The effect of haemolysis on the quantification and normalisation of miRNAs in plasma has not been investigated in great detail. We found that levels of miR-16, a commonly used reference gene, showed little variation when measured in plasma samples from healthy volunteers or patients with malignant mesothelioma or coronary artery disease. Including samples with evidence of haemolysis led to variation in miR-16 levels and consequently decreased its ability to serve as a reference. The levels of miR-16 and miR-451, both present in significant levels in red blood cells, were proportional to the degree of haemolysis. Measurements of the level of these miRNAs in whole blood, plasma, red blood cells and peripheral blood mononuclear cells revealed that the miRNA content of red blood cells represents the major source of variation in miR-16 and miR-451 levels measured in plasma. Adding lysed red blood cells to non-haemolysed plasma allowed a cut-off level of free haemoglobin to be determined, below which miR-16 and miR-451 levels displayed little variation between individuals. In conclusion, increases in plasma miR-16 and miR-451 are caused by haemolysis. In the absence of haemolysis the levels of both miR-16 and miR-451 are sufficiently constant to serve as normalisers.
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DOI：
10.1146/annurev.pathol.4.110807.092222
发表时间：
2009
期刊：
Annual review of pathology
影响因子：
--
作者：
Lee YS;Dutta A
通讯作者：
Dutta A
影响因子：
3.7
作者：
Collino F;Deregibus MC;Bruno S;Sterpone L;Aghemo G;Viltono L;Tetta C;Camussi G
通讯作者：
Camussi G
影响因子：
2.6
作者：
Choong, Meng Ling;Yang, Henry He;McNiece, Ian
通讯作者：
McNiece, Ian
影响因子：
9.3
作者：
Adachi, Taichi;Nakanishi, Michio;Iwai, Naoharu
通讯作者：
Iwai, Naoharu
影响因子：
6.5
作者：
Lawrie, Charles H.;Gal, Shira;Harris, Adrian L.
通讯作者：
Harris, Adrian L.