The Impact of Hemolysis on Cell-Free microRNA Biomarkers.
The Impact of Hemolysis on Cell-Free microRNA Biomarkers.
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DOI：
10.3389/fgene.2013.00094
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发表时间：
2013
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;Edelman JJ;Kao SC;Vallely MP;van Zandwijk N;Reid G
血浆和血清中的无细胞microRNA已成为各种疾病的生物标志物的有前途的来源。尽管在这一领域取得了快速进展，但仍然缺乏关于最佳定量方法，参考基因和样品质量控制的共识。最近的研究表明，在血液采集过程中发生的溶血对血浆/血清中的microRNA含量有实质性影响。迄今为止，仅对有限数量的microRNA研究了溶血的影响，主要是红细胞（RBC）富集的miR-16和-451。相比之下，溶血对其他microRNA的影响-特别是那些被提议作为生物标志物的microRNA-尚未得到解决。在这项研究中，我们分析了溶血和非溶血血浆以及RBC的microRNA含量，以获得受溶血影响或不受溶血影响的循环中microRNA的分布。使用TaqMan阵列微流体卡进行分析，以比较三对溶血和非溶血血浆（具有不同程度的溶血）和一份RBC样本。在至少两个样本中总共检测到136种microRNA，其中15种在所有三个溶血样本中至少升高了两倍。对于具有最高溶血水平的样品，该数量增加到88个microRNA，所有这些也在RBC谱中检测到。因此，这些microRNA代表了可检测microRNA的大部分，并且是最可能受溶血影响的microRNA。几种溶血敏感的microRNA（例如，miR-21、-106 a、-92 a、-17、-16）先前也已被提出作为疾病的血浆/血清生物标志物，突出了用于测量循环microRNA的血浆/血清样品的严格质量控制的重要性。由于低水平溶血在血浆/血清采集期间经常发生，因此在测量任何候选循环microRNA时考虑这一点至关重要。
Cell-free microRNAs in plasma and serum have become a promising source of biomarkers for various diseases. Despite rapid progress in this field, there remains a lack of consensus regarding optimal quantification methods, reference genes, and quality control of samples. Recent studies have shown that hemolysis occurring during blood collection has substantial impact on the microRNA content in plasma/serum. To date, the impact of hemolysis has only been investigated for a limited number of microRNAs, mainly the red blood cell (RBC)-enriched miRs-16 and -451. In contrast, the effect of hemolysis on other microRNAs – in particular those proposed as biomarkers – has not been addressed. In this study we profiled the microRNA content of hemolyzed and non-hemolyzed plasma as well as RBCs to obtain a profile of microRNAs in the circulation affected or unaffected by hemolysis. Profiling by TaqMan Array Microfluidic Cards was used to compare three pairs of hemolyzed and non-hemolyzed plasma (with varying degrees of hemolysis) and one RBC sample. A total of 136 microRNAs were detectable in at least two of the samples, and of those 15 were at least twofold elevated in all three hemolyzed samples. This number increased to 88 microRNAs for the sample with the highest level of hemolysis, with all of these also detected in the RBC profile. Thus these microRNAs represent a large proportion of detectable microRNAs and those most likely to be affected by hemolysis. Several of the hemolysis-susceptible microRNAs (e.g., miRs-21, -106a, -92a, -17, -16) have also been previously proposed as plasma/serum biomarkers of disease, highlighting the importance of rigorous quality control of plasma/serum samples used for measurement of circulating microRNAs. As low-level hemolysis is a frequent occurrence during plasma/serum collection it is critical that this is taken into account in the measurement of any candidate circulating microRNA.