title: Sensitive Surface Enhanced Raman Scattering-Based Detection of a BIGH3 Point Mutation Associated with Avellino Corneal Dystrophy.
abstract: Surface enhanced Raman scattering (SERS) is highly useful for sensitive analytical sensing; however, its practical availability for detecting a point mutation associated with disease in clinical sample was rarely proved. Herein, we present a toehold-mediated, DNA displacement-based, SERS sensor for detecting point mutations in the BIGH3 gene associated with the most common corneal dystrophies (CDs) in a clinical setting. To diagnose Avellino corneal dystrophy (ACD), selectivity was ensured by exploring optimal DNA displacement conditions such as length of toehold and hybridization temperature. A SERS-efficient Ag@Au bimetallic nanodendrite was employed to ensure sensitivity. Optimization for a clinical setting showed that discrimination was maximized when toehold length was 6-mer (T6), and hybridization temperature was 36 °C. On the basis of tests that used clinical homozygous and heterozygous CD samples, a single-base mismatched DNA sequence was identifiable within 30 min with a limit of detection (LOD) of 400 fM. From the results, we conclude that our toehold-mediated, DNA displacement-based, SERS sensor allows a rapid and sensitive detection of a BIGH3 gene point mutation associated with Avellino corneal dystrophy, indicating the practical ability of the method to diagnose genetic diseases caused by point mutations.
authors.affiliation: Department of Chemistry and Research Institute for Convergence of Basic Sciences, Hanyang University , Seoul, 04763, Republic of Korea.
authors.forename: Saetbyeol
authors.initials: S
authors.lastname: Kim
authors.affiliation: BIO-IT Foundry Technology Institute, Pusan National University , Busan, 46287, Republic of Korea.|Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital , Yangsan, Gyeongsangnam-do 50612, Republic of Korea.
authors.forename: Su-Nam
authors.initials: SN
authors.lastname: Jeong
authors.affiliation: Department of Chemistry and Research Institute for Convergence of Basic Sciences, Hanyang University , Seoul, 04763, Republic of Korea.
authors.forename: Sangsu
authors.initials: S
authors.lastname: Bae
authors.affiliation: Department of Chemistry and Research Institute for Convergence of Basic Sciences, Hanyang University , Seoul, 04763, Republic of Korea.
authors.forename: Hoeil
authors.initials: H
authors.lastname: Chung
authors.affiliation: BIO-IT Foundry Technology Institute, Pusan National University , Busan, 46287, Republic of Korea.|Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital , Yangsan, Gyeongsangnam-do 50612, Republic of Korea.
authors.forename: So Young
authors.initials: SY
authors.lastname: Yoo
mesh_terms: D003317:Corneal Dystrophies, Hereditary / Q000235:genetics*; D016326:Extracellular Matrix Proteins / Q000235:genetics*; D006801:Humans; D017354:Point Mutation; D013059:Spectrum Analysis, Raman; D013499:Surface Properties; D016212:Transforming Growth Factor beta / Q000235:genetics*; D000099222:betaIG-H3 Protein
chemical_list: D016326:Extracellular Matrix Proteins; D016212:Transforming Growth Factor beta; D000099222:betaIG-H3 Protein