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Anal Chem. 2016 Dec 6;88(23):11288-11292. Epub 2016 Nov 14.

Sensitive Surface Enhanced Raman Scattering-Based Detection of a BIGH3 Point Mutation Associated with Avellino Corneal Dystrophy.

Author information

1
Department of Chemistry and Research Institute for Convergence of Basic Sciences, Hanyang University , Seoul, 04763, Republic of Korea.
2
BIO-IT Foundry Technology Institute, Pusan National University , Busan, 46287, Republic of Korea.
3
Research Institute for Convergence of Biomedical Science and Technology, Pusan National University Yangsan Hospital , Yangsan, Gyeongsangnam-do 50612, Republic of Korea.

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.

PMID:
27934116
DOI:
10.1021/acs.analchem.6b03320
[Indexed for MEDLINE]

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