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Streptavidin-enhanced assay for sensitive and specific detection of single nucleotide polymorphism in TP53
H. Šípová, T. Špringer, J. Homola
Language English Country Germany
Document type Journal Article, Research Support, Non-U.S. Gov't
NLK
ProQuest Central
from 2011-01-01 to 1 year ago
Medline Complete (EBSCOhost)
from 2003-01-01 to 1 year ago
Health & Medicine (ProQuest)
from 2011-01-01 to 1 year ago
- MeSH
- Biosensing Techniques methods MeSH
- Molecular Diagnostic Techniques methods MeSH
- DNA Probes genetics MeSH
- DNA genetics MeSH
- Genes, p53 genetics MeSH
- Polymorphism, Single Nucleotide genetics MeSH
- Humans MeSH
- Oligonucleotides genetics MeSH
- Surface Plasmon Resonance instrumentation methods MeSH
- Sensitivity and Specificity MeSH
- Streptavidin chemistry MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Research Support, Non-U.S. Gov't MeSH
This paper reports an approach to detection of single nucleotide polymorphism based on special amplification assay and surface plasmon resonance biosensor technology. In this assay, a part of the target DNA is recognized by a probe (probe A) coupled with streptavidin-oligonucleotide (SON) complexes ex situ, and when the mixture is injected in the sensor, another part of the target DNA is recognized by a DNA probe (probe B) immobilized on the sensor surface. To achieve high sensitivity and specificity, the assay is optimized in terms of composition of SON complexes, probe design, and assay temperature. It is demonstrated that this approach provides high specificity (no response to targets containing single-mismatched bases) and sensitivity (improves sensor response to perfectly matched oligonucleotides by one order of magnitude compared to the direct detection method). The assay is applied to detection of a short synthetic analogue of TP53 containing a "hot spot"-single nucleotide mismatch frequently mutated in germ line cancer-at levels down to 40 pM.
References provided by Crossref.org
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