Quantitative analysis of microRNA in blood serum with protein-facilitated affinity capillary electrophoresis.
Anal Chem
; 83(16): 6196-201, 2011 Aug 15.
Article
en En
| MEDLINE
| ID: mdl-21714529
ABSTRACT
MicroRNAs (miRNAs) are small (â¼22 nt) regulatory RNAs that are frequently deregulated in cancer and have shown promise as tissue- and blood-based biomarkers for cancer classification and prognostication. Here we present a protein-facilitated affinity capillary electrophoresis (ProFACE) assay for rapid quantification of miRNA levels in blood serum using single-stranded DNA binding protein (SSB) and double-stranded RNA binding protein (p19) as separation enhancers. The method utilizes either the selective binding of SSB to a single-stranded DNA/RNA probe or the binding of p19 to miRNA-RNA probe duplex. For the detection of ultralow amounts of miRNA without polymerase chain reaction (PCR) amplification in blood samples we apply off-line preconcentration of synthetic miRNA-122 from serum by p19-coated magnetic beads followed by online sample stacking in the ProFACE assay. The detection limit is 0.5 fM or 30 000 miRNA molecules in 1 mL of serum as a potential source of naïve miRNAs.
Texto completo:
1
Colección:
01-internacional
Banco de datos:
MEDLINE
Asunto principal:
ADN de Cadena Simple
/
ARN Bicatenario
/
Técnicas de Química Analítica
/
Proteínas de Unión al ARN
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Electroforesis Capilar
/
MicroARNs
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Proteínas de Unión al ADN
Límite:
Humans
Idioma:
En
Revista:
Anal Chem
Año:
2011
Tipo del documento:
Article
País de afiliación:
Canadá