Improved Dried Blood Spot-Based Metabolomics: A Targeted, Broad-Spectrum, Single-Injection Method
Improved Dried Blood Spot-Based Metabolomics: A Targeted, Broad-Spectrum, Single-Injection Method
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DOI：
10.3390/metabo10030082
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发表时间：
2020-03-01
期刊：
影响因子：
4.1
通讯作者：
Naviaux, Robert K.
中科院分区：
文献类型：
--
作者：
Li, Kefeng;Naviaux, Jane C.;Naviaux, Robert K.
干血斑（DBS）已被证明是一种强大的采样和存储方法，用于新生儿筛查和许多其他应用。然而，DBS方法尚未优化用于广谱靶向代谢组学分析。在这项研究中，我们开发了一个强大的，基于DBS的，广谱的，有针对性的代谢物组学方法，能够测量超过400种代谢物，从6.3毫米打孔标准Whatman 903（TM）滤纸卡。分析了血斑体积、红细胞压积、真空采血管化学、提取方法、残留以及与血浆和手指针刺毛细血管血样的可比性的影响。还测试了在不同条件下储存超过一年的400种代谢物的稳定性。当点样血量超过60 μ L且红细胞压积在31%和50%之间时，未观察到血量和红细胞压积变化的显著影响。DBS代谢物组中代谢物的曲线下面积（AUC）中位数在前3个月下降了40%，然后至少1年没有进一步下降。所有最初可检测的代谢物均保持在可检测限度内。代谢组学分析的最佳储存条件是-80 ℃，干燥剂和无O-2清除剂。该方法被临床验证其在线粒体疾病线粒体脑肌病，乳酸酸中毒和中风样发作（MELAS）的诊断中的潜在效用。我们的方法为比较代谢组学研究提供了一种方便的冷冻、储存和运输液体血液样品的替代方法。
Dried blood spots (DBS) have proven to be a powerful sampling and storage method for newborn screening and many other applications. However, DBS methods have not yet been optimized for broad-spectrum targeted metabolomic analysis. In this study, we developed a robust, DBS-based, broad-spectrum, targeted metabolomic method that was able to measure over 400 metabolites from a 6.3 mm punch from standard Whatman 903(TM) filter paper cards. The effects of blood spot volumes, hematocrit, vacutainer chemistry, extraction methods, carryover, and comparability with plasma and fingerstick capillary blood samples were analyzed. The stability of over 400 metabolites stored under varying conditions over one year was also tested. No significant impacts of blood volume and hematocrit variations were observed when the spotted blood volume was over 60 mu L and the hematocrit was between 31% and 50%. The median area under the curve (AUC) of metabolites in the DBS metabolome declined by 40% in the first 3 months and then did not decline further for at least 1 year. All originally detectable metabolites remained within detectable limits. The optimal storage conditions for metabolomic analysis were -80 degrees C with desiccants and without an O-2 scavenger. The method was clinically validated for its potential utility in the diagnosis of the mitochondrial disease mitochondrial encephalomyopathy, lactic acidosis, and stroke-like episodes (MELAS). Our method provides a convenient alternative to freezing, storing, and shipping liquid blood samples for comparative metabolomic studies.