Polyethylene glycol improves current methods for circulating extracellular vesicle-derived DNA isolation
Polyethylene glycol improves current methods for circulating extracellular vesicle-derived DNA isolation
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DOI：
10.1186/s12967-019-1825-3
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发表时间：
2019-03-11
影响因子：
7.4
通讯作者：
Ayuso-Sacido, A.
中科院分区：
文献类型：
--
作者：
Garcia-Romero, N.;Madurga, R.;Ayuso-Sacido, A.
细胞外小泡是一种小的膜结合小泡，在细胞间的通讯中起着重要的作用。它们的分子载物分析是生物标志物检测的新来源，它可能提供传统固体活检的替代方案。结果我们研究了最常见的EV分离方法--超速离心法、聚乙二醇法和两种商用试剂盒ExoQuick((R))和PureExo((R))的效率。我们从健康供者的血液中分离出循环中的EVS，表征了EVS的大小和产量，并分析了它们的蛋白质谱和浓度。此外，我们首次使用Digital-PCR来鉴定和检测特定的gDNA序列，这对多种疾病的诊断和监测具有一定的意义。结论聚乙二醇沉淀法是最可行且成本更低的EV分离技术。
BackgroundExtracellular vesicles (EVs) are small membrane-bound vesicles which play an important role in cell-to-cell communication. Their molecular cargo analysis is presented as a new source for biomarker detection, and it might provide an alternative to traditional solid biopsies. However, the most effective approach for EV isolation is not yet wellestablished.ResultsHere, we study the efficiency of the most common EV isolation methods-ultracentrifugation, Polyethlyene glycol and two commercial kits, Exoquick((R)) and PureExo((R)). We isolated circulating EVs from the bloodstream of healthy donors, characterized the size and yield of EVs and analyzed their protein profiles and concentration. Moreover, we have used for the first time Digital-PCR to identify and detect specific gDNA sequences, which has several implications for diagnostic and monitoring many types of diseases.ConclusionsOur findings present Polyethylene glycol precipitation as the most feasible and less cost-consuming EV isolation technique.