Optical detection and discrimination of cystic fibrosis-related genetic mutations using oligonucleotide-nanoparticle conjugates.
Novel methods for application of oligonucleotide-gold nanoparticle conjugates to selective colorimetric detection and discrimination of cystic fibrosis (CF) related genetic mutations in model oligonucleotide systems are presented. Three-strand oligonucleotide complexes are employed, wherein two probe oligonucleotide-gold nanoparticle conjugates are linked together by a third target oligonucleotide strand bearing the CF-related mutation(s). By monitoring the temperature dependence of the optical properties of the complexes, either in solution or on silica gel plates, melting behaviors may be accurately and reproducibly compared. Using this approach, fully complementary sequences are successfully distinguished from mismatched sequences, with single base mismatch resolution, for Delta F 508, M470V, R74W and R75Q mutations.
Murphy
Deirdre
D
Nanotechnology Group, Tyndall National Institute, Lee Maltings, Cork, Ireland.
Redmond
Gareth
G
D020137:Base Pair Mismatch; D001483:Base Sequence; D001798:Blood Proteins / Q000235:genetics*; D003550:Cystic Fibrosis / Q000235:genetics* / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D006046:Gold; D006801:Humans; D009154:Mutation; D036103:Nanotechnology; D009841:Oligonucleotides / Q000737:chemistry / Q000235:genetics*; D010316:Particle Size; D058428:Silica Gel; D012822:Silicon Dioxide / Q000737:chemistry; D012996:Solutions / Q000737:chemistry; D013696:Temperature
D001798:Blood Proteins; C505032:CFTR protein, human; D009841:Oligonucleotides; D012996:Solutions; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D058428:Silica Gel; D006046:Gold; D012822:Silicon Dioxide