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Multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing corneal dystrophy, and method for producing same

A nanostructure and nucleic acid chip technology, applied to array nucleic acid chips, and its preparation method can solve problems such as difficult simultaneous measurement and long manufacturing time

Inactive Publication Date: 2012-08-22
KOREA ADVANCED INST OF SCI & TECH +1
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such LSPR-based biosensors still have the problems of long manufacturing time and difficulty in simultaneously measuring and discriminating multiple analyte biomolecules

Method used

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  • Multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing corneal dystrophy, and method for producing same
  • Multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing corneal dystrophy, and method for producing same
  • Multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing corneal dystrophy, and method for producing same

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0063] Example 1: Preparation of multi-point metal-capped nanostructure array nucleic acid chip for diagnosis of corneal dystrophy

[0064] Using a vacuum deposition system (Shinu MST Co., Ltd., Korea), each of chromium and gold was vacuum-deposited on the surface of a glass slide substrate (first layer, 75 mm × 25 mm × 1 mm), thereby forming a gold thin film layer (second Floor). Specifically, chromium is deposited as an intermediate metal thin film to a thickness of 5 nm, and the thickness of the gold thin film layer can be controlled to be 40 nm. Then, the porous mask (15, 20, 60 or 140 points) was immobilized onto the gold thin film layer (second layer) by adsorption. Then, on the surface of the gold thin film layer (second layer) on which the porous mask was adsorbed, a SAM film was formed using 1 mM 4,4'-dithiodibutyric acid (DDA). Then 400 mM EDC was added to the surface of the gold thin film layer (second layer), and activation was performed on the carboxyl groups ...

Embodiment 2

[0065] Example 2: Construction of Nanostructured Array Nucleic Acid Chips Including Multipoint Metal Capping for Diagnosis of Corneal Dystrophy Without LSPR Optical Characteristic Labeling

[0066] figure 2 A photograph of an optical biodetector without LSPR-based optical property labeling comprising a multipoint metal-terminated nanostructure array nucleic acid chip prepared as described above is shown. Optical biodetectors including tungsten-halogen light sources (wavelength: 360nm-2000nm, Ocean Optics, Inc., USA), detectors (wavelength: 300nm-1100nm, Ocean Optics , Inc., USA), a spectrophotometer for separating the light detected by the detector (wavelength: 200nm-1100nm, Ocean Optics, Inc., USA), and analysis / processing for processing the results obtained in the spectrophotometer program (Ocean Optics, Inc., USA). Here, the tungsten-halogen light source and detector are included in one light source probe. The incident light emitted from the optical biodetector withou...

Embodiment 3

[0067] Example 3: Determination of BIGH3 gene mutation type

[0068] In order to construct a probe for diagnosing mutations in the BIGH3 gene responsible for eye diseases, including Avellino corneal dystrophy, the BIGH3 gene mutation site for construction of the probe was determined. The DNA base sequence and amino acid sequence of the BIGH3 gene mutation site were analyzed and obtained through NCBI gene-related database GenBank and OMIM (Online Mendelian Human Genetics Database), and the information of each allele was also obtained. In order to test the effectiveness of the diagnostic chip, the type of mutation to be searched is first determined. Among the BIGH3 gene hotspots, the mutations causing Avellino corneal dystrophy (ACD), lattice corneal dystrophy type Ⅰ (LCD) and Reis-bucklers corneal dystrophy (RBCD) were selected (Table 1). Mutations in the exon 4 region.

[0069] Table 1. Eye diseases with BIGH3 gene mutation

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Abstract

The present invention relates to a multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing corneal dystrophy, and more particularly, to a multi-spot metal-deposited nucleic acid chip with nanostructure arrays which uses the optical characteristics of localized surface plasmon resonance (LSPR), to a method for producing the chip, and to a multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing BIGH3 gene mutations and various types of corneal dystrophies. According to the present invention, the metal-deposited nucleic acid chip with nanostructure arrays, and an analysis device including a light source, a detector, a spectrophotometer and a computer are combined to be used as a label-free biosensor based on the optical characteristics of LSPR. The multi-spot metal-deposited nucleic acid chip with nanostructure arrays for diagnosing BIGH3 gene mutations and corneal dystrophy according to the present invention diagnoses various types of corneal dystrophies, which are hereditary eye diseases, at the same time.

Description

technical field [0001] The present invention relates to a multi-point metal-terminated nanostructure array nucleic acid chip for diagnosing corneal dystrophy, more particularly, the present invention relates to a multi-point metal-terminated nanostructure array capable of employing LSPR (localized surface plasmon resonance) optical properties A nucleic acid chip, its preparation method, and a multi-point metal-capped nanostructure array nucleic acid chip for diagnosing BIGH3 gene mutations, the chip can diagnose various corneal dystrophies. Background technique [0002] Corneal dystrophy is an autosomal dominant genetic disorder in which a shadow develops in the center of the cornea and increases with age, reducing vision. Corneal dystrophies include Avellino corneal dystrophy (ACD), lattice corneal dystrophy type I (LCD), and Reis-bucklers corneal dystrophy (RBCD), and are caused by mutations in the gene encoding the BIGH3 protein. Patients with heterozygous Avellino corne...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12Q1/68
CPCB01J2219/00509B01J2219/00608B01J2219/00612B01J2219/00614B01J2219/00621B01J2219/00637B01J2219/00648B01J2219/00722B82Y30/00C12Q1/6883C12Q2600/158C12Q1/6837C12Q2563/155
Inventor 李相烨刘小永金道钧朴泰桢尹正国李津
Owner KOREA ADVANCED INST OF SCI & TECH
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