Visual, base-specific detection of nucleic acid hybridization using polymerization-based amplification

Polymerization-based signal amplification offers sensitive visualization of biotinylated biomolecules functionalized to glass microarrays in a manner suitable for point-of-care use. Here we report using this method for visual detection of multiplexed nucleic acid hybridizations from complex media an...

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Bibliographic Details
Published in:Analytical biochemistry 2009-03, Vol.386 (2), p.285-287
Main Authors: Hansen, Ryan R., Johnson, Leah M., Bowman, Christopher N.
Format: Article
Language:English
Subjects:
Codon
DNA - analysis
DNA - genetics
Humans
Nucleic Acid Amplification Techniques - instrumentation
Nucleic Acid Amplification Techniques - methods
Nucleic Acid Hybridization - methods
Polymers - chemistry
Polymorphism, Single Nucleotide
Proto-Oncogene Proteins - genetics
Proto-Oncogene Proteins p21(ras)
ras Proteins - genetics
Sensitivity and Specificity
Tumor Suppressor Protein p53 - genetics
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Summary:Polymerization-based signal amplification offers sensitive visualization of biotinylated biomolecules functionalized to glass microarrays in a manner suitable for point-of-care use. Here we report using this method for visual detection of multiplexed nucleic acid hybridizations from complex media and develop an application toward point mutation detection and single nucleotide polymorphism (SNP) typing. Primer extension reactions were employed to label selectively and universally all complementary surface DNA hybrids with photoinitiators, permitting simultaneous and dynamic photopolymerization from positive sites to 0.5-nM target concentrations. Dramatic improvements in signal ratios between complementary and mismatched hybrids enabled visual discrimination of single base differences in KRAS codon-12 biomarkers.
ISSN:0003-2697
1096-0309
DOI:10.1016/j.ab.2008.12.009