Development of diagnostic platforms for single nucleotide polymorphism detection

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2014
Akça, Oya
In this study, we report the results of different simple and cost effective approaches and platforms which were used to improve single nucleotide mismatch discrimination ability and investigation of the applicability of various materials and environments for the system. As the solid support of the array platform both customly functionalized epoxy and amine combination and commercially purchased poly-L-lysine modified materials were used. Surface-probe interactions and immobilization efficiencies were investigated and affirmative results are gained with heterobifunctional cross linker mediated immobilization of polyethyleneglycol spacer functionalized surface probes. Glass capillary tubes were also used to develop a partially solution based approach that can detect single nucleotide polymorphisms (SNPs) through the Oligonucleotide Ligation Reaction. The established sytem was capable of discriminating mistmatches at single nucleotide level by using unlabeled targets. Mesoporous silica nanoparticles and nitrocellulose membrane strips were also used to develop a novel platform for SNP detection. Controlled release of chromogen molecules inside the pores of the silica nanoparticles were achieved related with the specific complementary probe hybridization events. Use of unmodified target sequences in sandwich hybridization format permitted the usage of different detection signal probes throughout the study. Signal probes modified with 6-carboxyfluorescein or biotin at 3’ ends were successfully used to visualize the experimental results by using simple UV illumination and of color formation. This enables the platform to be followed by unaided eyes which makes it an ideal candidate for point of care detection purposes.

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Citation Formats
O. Akça, “Development of diagnostic platforms for single nucleotide polymorphism detection,” Ph.D. - Doctoral Program, Middle East Technical University, 2014.