{"title":"[MicroRNA分析血浆制备标准化:高速离心和膜过滤的影响]。","authors":"Saki Shiraihama, Yuki Kobayashi, Chieri Kuroki, Mayuko Hirotsu, Kazuhiro Okada, Tatsuki Shibuta, Hiromichi Shiotsu, Taeko Hotta, Dongchon Kang, 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":"65 3","pages":"260-265"},"PeriodicalIF":0.0000,"publicationDate":"2017-03-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"[Standardization of Plasma Preparation for MicroRNA Analysis: Influences of High Speed Centrifugation and Membrane Filtration].\",\"authors\":\"Saki Shiraihama, Yuki Kobayashi, Chieri Kuroki, Mayuko Hirotsu, Kazuhiro Okada, Tatsuki Shibuta, Hiromichi Shiotsu, Taeko Hotta, Dongchon Kang, 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\":\"65 3\",\"pages\":\"260-265\"},\"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}","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}
[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].