Analytics of Cerebrospinal Fluid MicroRNA Quantitative PCR Studies.
Analytics
Cerebrospinal fluid
Quantitative PCR
TaqMan low-density array
microRNA
Journal
Molecular neurobiology
ISSN: 1559-1182
Titre abrégé: Mol Neurobiol
Pays: United States
ID NLM: 8900963
Informations de publication
Date de publication:
Jul 2019
Jul 2019
Historique:
received:
14
08
2018
accepted:
02
11
2018
pubmed:
16
11
2018
medline:
15
1
2020
entrez:
16
11
2018
Statut:
ppublish
Résumé
MicroRNAs (miRNAs) are small non-coding RNAs that regulate post-transcriptional gene expression. Recent studies have shown that human disease states correlate with measurable differences in the level of circulating miRNAs relative to healthy controls. Thus, there is great interest in developing clinical miRNA assays as diagnostic or prognostic biomarkers for diseases, and as surrogate measures for therapeutic outcomes. Our studies have focused on miRNAs in human cerebral spinal fluid (CSF) as biomarkers for central nervous system (CNS) diseases. Our objective here was to examine factors that may affect the outcome of quantitative PCR (qPCR) studies on CSF miRNAs, in order to guide planning and interpretation of future CSF miRNA TaqMan® low-density array (TLDA) studies. We obtained CSF from neurologically normal (control) donors and used TLDAs to measure miRNA expression. We examined sources of error in the TLDA outcomes due to (1) nonspecific amplification of products in total RNA, (2) variations in RNA isolations performed on different days, (3) miRNA primer probe efficiency, and (4) variations in individual TLDA cards. We also examined the utility of card-to-card TLDA corrections and use of an unchanged "reference standard" to remove batch processing effects in large-scale studies.
Identifiants
pubmed: 30430409
doi: 10.1007/s12035-018-1422-0
pii: 10.1007/s12035-018-1422-0
pmc: PMC6527498
mid: NIHMS1524646
doi:
Substances chimiques
Biomarkers
0
MicroRNAs
0
Types de publication
Journal Article
Langues
eng
Sous-ensembles de citation
IM
Pagination
4988-4999Subventions
Organisme : NCATS NIH HHS
ID : UL1 TR000128
Pays : United States
Organisme : NCATS NIH HHS
ID : UL1 TR002369
Pays : United States
Organisme : NIA NIH HHS
ID : P30AG008017
Pays : United States
Organisme : NCATS NIH HHS
ID : UH2 TR000903
Pays : United States
Organisme : NIA NIH HHS
ID : P30 AG008017
Pays : United States
Organisme : NCATS NIH HHS
ID : Oregon Clinical & Translational Research Institute UL1TR000128
Pays : United States
Organisme : NCATS NIH HHS
ID : UH3 TR000903
Pays : United States
Organisme : NCATS NIH HHS
ID : UH2/UH3TR000903
Pays : United States
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