[MicroRNA分析血浆制备标准化:高速离心和膜过滤的影响]。

Saki Shiraihama, Yuki Kobayashi, Chieri Kuroki, Mayuko Hirotsu, Kazuhiro Okada, Tatsuki Shibuta, Hiromichi Shiotsu, Taeko Hotta, Dongchon Kang, Tsukuru Umemura
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引用次数: 0

摘要

MicroRNAs (miRNA)是非编码小rna。外泌体在血浆和其他体液中携带细胞外mirna。血浆mirna水平显示疾病特异性变化。因此,mirna有望成为许多疾病的新生物标志物。然而,血浆mirna的分析方法尚不完善。在这项研究中,我们使用基于逆转录酶的定量聚合酶链反应(RT-qPCR)测试了高速离心和膜过滤对血浆miRNA分析结果的影响。我们研究了12名正常人的血浆。经高速离心过滤后血浆miR-451水平无明显变化,略有升高,为1.543±0.263倍(mean±SD, N=3)。经高速离心和过滤后,血浆miR-126和miR-223水平分别下降0.038±0.008倍和0.041±0.003倍。我们的数据表明,通过高速离心和/或过滤从血浆中去除血小板和细胞碎片对于血浆miRNA分析的标准化至关重要。(最初的)。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
[Standardization of Plasma Preparation for MicroRNA Analysis: Influences of High Speed Centrifugation and Membrane Filtration].

MicroRNAs (miRNA) are non-coding small RNAs. Exosomes carry extracellular miRNAs in plasma and other body fluids. Levels of plasma miRNAs show disease-specific changes. Thus, miRNAs are expected to be new biomarkers in many diseases. However, the method of analysis of plasma miRNAs is not well established. In this study, we tested the influences of high speed centrifugation and membrane filtration on results from plasma miRNA analysis using reverse transcriptase-based quantitative polymerase chain reac- tion (RT-qPCR). We studied plasma from 12 normal subjects. The level of plasma miR-451 did not change significantly after high speed centrifugation and filtration, rather showed slight increment, 1.543 ± 0.263 fold (mean±SD, N=3). The levels of plasma miR-126 and miR-223 decreased with high speed centrifugation and filtration, (0.038 ± 0.008 fold and 0.041 ± 0.003 fold, respectively). Our data suggested that removing platelets and cellular debris from plasma with high speed centrifugation and/or filtration is essential for stand- ardization of plasma miRNA analysis. [Original].

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