Detail
Article
Online article
FT
Medvik - BMC
  • Something wrong with this record ?

Two-tailed RT-qPCR panel for quality control of circulating microRNA studies

P. Androvic, N. Romanyuk, L. Urdzikova-Machova, E. Rohlova, M. Kubista, L. Valihrach,

. 2019 ; 9 (1) : 4255. [pub] 20190312

Language English Country Great Britain

Document type Journal Article, Research Support, Non-U.S. Gov't

Persistent link   https://www.medvik.cz/link/bmc20028937

Circulating cell-free microRNAs are promising candidates for minimally invasive clinical biomarkers for the diagnosis, prognosis and monitoring of many human diseases. Despite substantial efforts invested in the field, the research so far has failed to deliver expected results. One of the contributing factors is general lack of agreement between various studies, partly due to the considerable technical challenges accompanying the workflow. Pre-analytical variables including sample collection, RNA isolation, and quantification are sources of bias that may hamper biological interpretation of the results. Here, we present a Two-tailed RT-qPCR panel for quality control, monitoring of technical performance, and optimization of microRNA profiling experiments from biofluid samples. The Two-tailed QC (quality control) panel is based on two sets of synthetic spike-in molecules and three endogenous microRNAs that are quantified with the highly specific Two-tailed RT-qPCR technology. The QC panel is a cost-effective way to assess quality of isolated microRNA, degree of inhibition, and erythrocyte contamination to ensure technical soundness of the obtained results. We provide assay sequences, detailed experimental protocol and guide to data interpretation. The application of the QC panel is demonstrated on the optimization of RNA isolation from biofluids with the miRNeasy Serum/Plasma Advanced Kit (Qiagen).

References provided by Crossref.org

000      
00000naa a2200000 a 4500
001      
bmc20028937
003      
CZ-PrNML
005      
20210114155344.0
007      
ta
008      
210105s2019 xxk f 000 0|eng||
009      
AR
024    7_
$a 10.1038/s41598-019-40513-w $2 doi
035    __
$a (PubMed)30862831
040    __
$a ABA008 $b cze $d ABA008 $e AACR2
041    0_
$a eng
044    __
$a xxk
100    1_
$a Androvic, Peter $u Institute of Biotechnology of the Czech Academy of Sciences - BIOCEV, Vestec, 252 50, Czech Republic. Laboratory of Growth Regulators, Faculty of Science, Palacky University, Olomouc, 78371, Czech Republic.
245    10
$a Two-tailed RT-qPCR panel for quality control of circulating microRNA studies / $c P. Androvic, N. Romanyuk, L. Urdzikova-Machova, E. Rohlova, M. Kubista, L. Valihrach,
520    9_
$a Circulating cell-free microRNAs are promising candidates for minimally invasive clinical biomarkers for the diagnosis, prognosis and monitoring of many human diseases. Despite substantial efforts invested in the field, the research so far has failed to deliver expected results. One of the contributing factors is general lack of agreement between various studies, partly due to the considerable technical challenges accompanying the workflow. Pre-analytical variables including sample collection, RNA isolation, and quantification are sources of bias that may hamper biological interpretation of the results. Here, we present a Two-tailed RT-qPCR panel for quality control, monitoring of technical performance, and optimization of microRNA profiling experiments from biofluid samples. The Two-tailed QC (quality control) panel is based on two sets of synthetic spike-in molecules and three endogenous microRNAs that are quantified with the highly specific Two-tailed RT-qPCR technology. The QC panel is a cost-effective way to assess quality of isolated microRNA, degree of inhibition, and erythrocyte contamination to ensure technical soundness of the obtained results. We provide assay sequences, detailed experimental protocol and guide to data interpretation. The application of the QC panel is demonstrated on the optimization of RNA isolation from biofluids with the miRNeasy Serum/Plasma Advanced Kit (Qiagen).
650    _2
$a zvířata $7 D000818
650    _2
$a biologické markery $x krev $7 D015415
650    _2
$a cirkulující mikroRNA $x krev $x izolace a purifikace $7 D000074166
650    _2
$a analýza nákladů a výnosů $7 D003362
650    _2
$a studie proveditelnosti $7 D005240
650    _2
$a zdraví dobrovolníci pro lékařské studie $7 D064368
650    _2
$a lidé $7 D006801
650    12
$a řízení kvality $7 D011786
650    _2
$a krysa rodu Rattus $7 D051381
650    _2
$a reagenční diagnostické soupravy $x normy $7 D011933
650    _2
$a kvantitativní polymerázová řetězová reakce $x ekonomika $x přístrojové vybavení $x metody $x normy $7 D060888
655    _2
$a časopisecké články $7 D016428
655    _2
$a práce podpořená grantem $7 D013485
700    1_
$a Romanyuk, Nataliya $u Institute of Experimental Medicine of the Czech Academy of Sciences, Prague, 142 20, Czech Republic.
700    1_
$a Urdzikova-Machova, Lucia $u Institute of Experimental Medicine of the Czech Academy of Sciences, Prague, 142 20, Czech Republic.
700    1_
$a Rohlova, Eva $u Institute of Biotechnology of the Czech Academy of Sciences - BIOCEV, Vestec, 252 50, Czech Republic. Department of Anthropology and Human Genetics, Faculty of Science, Charles University, Prague, 128 43, Czech Republic.
700    1_
$a Kubista, Mikael $u Institute of Biotechnology of the Czech Academy of Sciences - BIOCEV, Vestec, 252 50, Czech Republic. TATAA Biocenter AB, Gothenburg, 411 03, Sweden.
700    1_
$a Valihrach, Lukas $u Institute of Biotechnology of the Czech Academy of Sciences - BIOCEV, Vestec, 252 50, Czech Republic. lukas.valihrach@ibt.cas.cz.
773    0_
$w MED00182195 $t Scientific reports $x 2045-2322 $g Roč. 9, č. 1 (2019), s. 4255
856    41
$u https://pubmed.ncbi.nlm.nih.gov/30862831 $y Pubmed
910    __
$a ABA008 $b sig $c sign $y a $z 0
990    __
$a 20210105 $b ABA008
991    __
$a 20210114155343 $b ABA008
999    __
$a ok $b bmc $g 1609272 $s 1120117
BAS    __
$a 3
BAS    __
$a PreBMC
BMC    __
$a 2019 $b 9 $c 1 $d 4255 $e 20190312 $i 2045-2322 $m Scientific reports $n Sci Rep $x MED00182195
LZP    __
$a Pubmed-20210105

Find record

Citation metrics

Archiving options