Development of a gold nano-particle-based fluorescent molecular beacon for detection of cystic fibrosis associated mutation.
Cystic fibrosis is one of the most common genetically inherited diseases in Northern Europe, consisting of an inherited defect of chloride transport in the epithelium. Of the several mutations related to CF, the ΔF508 mutation occurs in ca. 70% of the cases. In this work the use of a gold nano-particle supported fluorescence molecular beacon was investigated as an optical sensing platform for the detection of the ΔF508 cystic fibrosis associated mutation. Different parameters such as molecular beacon design, Au nano-particle size, molecular beacon-nano-particle conjugation protocol, molecular beacon loading as well as experimental conditions were evaluated. A 31-base long molecular beacon, containing a 15-base recognition sequence specific for the mutant target, was linked via a thiol modified poly thymine linker (10 bases long) to a 13 nm gold nano-particle and was exposed to mutant and wild type targets, and a clear differentiation was achieved at target concentrations as low as 1 nM.
Beni
Valerio
V
Nanobiotechnology & Bioanalysis Group, Departament d'Enginyeria Quimica, Universitat Rovira i Virgili, Avinguda Països Catalans, 26, 43007 Tarragona, Spain. valerio.beni@urv.cat
Hayes
Karen
K
Lerga
Teresa Mairal
TM
O'Sullivan
Ciara K
CK
D003550:Cystic Fibrosis / Q000175:diagnosis* / Q000235:genetics*; D004252:DNA Mutational Analysis / Q000295:instrumentation*; D004867:Equipment Design; D019544:Equipment Failure Analysis; D006046:Gold; D006801:Humans; D015336:Molecular Probe Techniques / Q000295:instrumentation*; D053758:Nanoparticles; D013050:Spectrometry, Fluorescence / Q000295:instrumentation*
D006046:Gold