Cystic Fibrosis Mutation Detection with SPR Biosensor in Real Samples via Multiple Surfaces Binding Method.

Cystic Fibrosis Mutation Detection with SPR Biosensor in Real Samples via Multiple Surfaces Binding Method. Comb Chem High Throughput Screen. 2017 Jan 03; Authors: Kayran YU, Ozkan-Ariksoysal D, Topkaya SN, Kaymaz BT, Can BK Abstract Surface Plasmon Resonance (SPR) based biosensor system was developed for the detection of Delta F508 (DF508del) Cystic Fibrosis (CF) mutation in both synthetic and real samples. In order to provide an effective hybridization between probe and the Polymerase Chain Reaction (PCR) amplicons (target), streptavidin was bound to the surface and biotin-tag probe was sent to the streptavidin-coated surface. For the target preparation, blood samples were collected from the patients who suffer from CF. Following the DNA isolation; samples were ampliļ¬ed with PCR with biotin-tag. Before sending the biotin-tag PCR amplicons onto the modified surface, amplicons were also interacted with the helper oligonucleotides to prevent re-annealing of the denatured DNA strands. This kind of 'multiple surface binding' method helps increasing of the sensitivity of the detection. The limit of detection(S/N= 3) was calculated as 12.24 pico-mole/ml for PCR-like synthetic long target sequence and 13x105 molecules for real samples in less than half hour. Using the both biotin-tag probe and the helper oligonucleotides together, hybridization was obtained much more efficiently than traditional denaturation protocols for real samples an...
Source: Combinatorial Chemistry and High Throughput Screening - Category: Chemistry Authors: Tags: Comb Chem High Throughput Screen Source Type: research