[Standardization of Plasma Preparation for MicroRNA Analysis: Influences of High Speed Centrifugation and Membrane Filtration].

Saki Shiraihama, Yuki Kobayashi, Chieri Kuroki, Mayuko Hirotsu, Kazuhiro Okada, Tatsuki Shibuta, Hiromichi Shiotsu, Taeko Hotta, Dongchon Kang, Tsukuru Umemura
{"title":"[Standardization of Plasma Preparation for MicroRNA Analysis: Influences of High Speed Centrifugation and Membrane Filtration].","authors":"Saki Shiraihama,&nbsp;Yuki Kobayashi,&nbsp;Chieri Kuroki,&nbsp;Mayuko Hirotsu,&nbsp;Kazuhiro Okada,&nbsp;Tatsuki Shibuta,&nbsp;Hiromichi Shiotsu,&nbsp;Taeko Hotta,&nbsp;Dongchon Kang,&nbsp;Tsukuru Umemura","doi":"","DOIUrl":null,"url":null,"abstract":"<p><p>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].</p>","PeriodicalId":21457,"journal":{"name":"Rinsho byori. The Japanese journal of clinical pathology","volume":null,"pages":null},"PeriodicalIF":0.0000,"publicationDate":"2017-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Rinsho byori. The Japanese journal of clinical pathology","FirstCategoryId":"1085","ListUrlMain":"","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"","JCRName":"","Score":null,"Total":0}
引用次数: 0

Abstract

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].

[MicroRNA分析血浆制备标准化:高速离心和膜过滤的影响]。
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。
求助全文
约1分钟内获得全文 求助全文
来源期刊
自引率
0.00%
发文量
0
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信