Detection of single-nucleotide polymorphisms using gold nanoparticles and single-strand-specific nucleases.
The current study reports an assay approach that can detect single-nucleotide polymorphisms (SNPs) and identify the position of the point mutation through a single-strand-specific nuclease reaction and a gold nanoparticle assembly. The assay can be implemented via three steps: a single-strand-specific nuclease reaction that allows the enzyme to truncate the mutant DNA; a purification step that uses capture probe-gold nanoparticles and centrifugation; and a hybridization reaction that induces detector probe-gold nanoparticles, capture probe-gold nanoparticles, and the target DNA to form large DNA-linked three-dimensional aggregates of gold nanoparticles. At high temperature (63 degrees C in the current case), the purple color of the perfect match solution would not change to red, whereas a mismatched solution becomes red as the assembled gold nanoparticles separate. Using melting analysis, the position of the point mutation could be identified. This assay provides a convenient colorimetric detection that enables point mutation identification without the need for expensive mass spectrometry. To our knowledge, this is the first report concerning SNP detection based on a single-strand-specific nuclease reaction and a gold nanoparticle assembly.
Chen
Yen-Ting
YT
Institute of Biotechnology, National Taipei University of Technology, Taipei 106, Taiwan.
Hsu
Chiao-Ling
CL
Hou
Shao-Yi
SY
D001483:Base Sequence; D003365:Costs and Cost Analysis; D004252:DNA Mutational Analysis / Q000295:instrumentation* / Q000379:methods*; D015342:DNA Probes / Q000378:metabolism; D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism*; D003851:Deoxyribonucleases / Q000378:metabolism*; D006046:Gold / Q000737:chemistry*; D053768:Metal Nanoparticles / Q000737:chemistry*; D009841:Oligonucleotides / Q000235:genetics / Q000378:metabolism; D017354:Point Mutation; D020641:Polymorphism, Single Nucleotide; D013379:Substrate Specificity; D013997:Time Factors; D044366:Transition Temperature
D015342:DNA Probes; D004277:DNA, Single-Stranded; D009841:Oligonucleotides; D006046:Gold; D003851:Deoxyribonucleases