Surface plasmon resonance imaging (SPRI) system and real-time monitoring of DNA biochip for human genetic mutation diagnosis of DNA amplified samples

摘要

Nucleic acid-based biochips represent a promising tool for gene sequence analysis, especially for mutation detection. Surface plasmon resonance imaging (SPRI) technique allows kinetic monitoring of molecular interaction, such as DNA-DNA, in real-time and without any prior labelling step. SPRI was applied to developing a DNA sensor for the detection of gene mutations accounting for human cystic fibrosis and specifically some of those localized in exon 10, with the common three-base-pair deletion ΔF508, alongside several single nucleotide polymorphisms (SNPs). The SPRI system enabled us to monitor the hybridisation kinetics of unlabelled DNA targets (short oligonucleotides or a 377 pb PCR product) to a 196 spots matrix of ssDNA probes immobilised onto a bio-functionalised surface, and to detect in real-time the mutations in a DNA fragment.