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 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.
Hansen
Ryan R
RR
Department of Chemical and Biological Engineering, University of Colorado, ECCH 111 CB 424, Boulder, CO 80309, USA.
Johnson
Leah M
LM
Bowman
Christopher N
CN
D003062:Codon; D004247:DNA / Q000032:analysis / Q000235:genetics; D006801:Humans; D021141:Nucleic Acid Amplification Techniques / Q000295:instrumentation / Q000379:methods*; D009693:Nucleic Acid Hybridization / Q000379:methods*; D011108:Polymers / Q000737:chemistry*; D020641:Polymorphism, Single Nucleotide; D011518:Proto-Oncogene Proteins / Q000235:genetics; D016283:Proto-Oncogene Proteins p21(ras); D012680:Sensitivity and Specificity; D016159:Tumor Suppressor Protein p53 / Q000235:genetics; D018631:ras Proteins / Q000235:genetics
D003062:Codon; C117307:KRAS protein, human; D011108:Polymers; D011518:Proto-Oncogene Proteins; D016159:Tumor Suppressor Protein p53; D004247:DNA; D016283:Proto-Oncogene Proteins p21(ras); D018631:ras Proteins