Detection of the Most Common Corneal Dystrophies Caused by BIGH3 Gene Point Mutations Using a Multispot Gold-Capped Nanoparticle Array Chip

摘要

The localized surface plasmon resonance (LSPR) opticalnproperty has recently been well employed as an effectivenplatform for the quantitative detection of protein-proteinninteractions on the nanoscale. However, its advantage hasnnot been fully explored yet in the DNA diagnosis field,nespecially in detecting point mutations of DNA. Pointnmutations of the BIGH3 gene are associated with the mostncommon corneal dystrophies (CDs), such as Avellinoncorneal dystrophy, Reis-Bucklers corneal dystrophy, andnlattice corneal dystrophy. Since the detection of thesencorneal dystrophies is urgently needed before laserassistednin situ keratomileusis operation to prevent blindness,ngenetic analysis of the BIGH3 gene is critical in mostnophthalmological clinics. In this study, we report LSPRbasedndetection of the BIGH3 gene mutations by using anmultispot gold-capped nanoparticle array (MG-NPA) chip.nThe analytical range and sensitivity of the MG-NPA chipnwere determined by measuring different concentrationsnof each CD target DNA in the range of 1 fMto 1 μM. Undernthe optimal conditions, the detection of DNA hybridizationnwith each CD target DNA was performed with a detectionnlimit of 1 pM target DNA. The selective discriminationnagainst a single-base mismatch DNA sequence was alsonachieved by using both homozygous and heterozygous CDnsamples. It demonstrates that the label-free LSPR-basednoptical biosensor system employing the MG-NPA chipnprovides a new diagnostic platform allowing the selectivenand sensitive detection of various DNA point mutations,nleading to possible diagnosis of mutation-related diseasesnincluding corneal dystrophies reported here.