Label-Free Fluorescent Copper Nanoclusters for Genotyping of Deletion and Duplication of Duchenne Muscular Dystrophy

Chung-An Chen, Chun Chi Wang, Yuh-Jyh Jong, Shou-Mei Wu

Research output: Contribution to journalArticlepeer-review

20 Scopus citations

Abstract

Real applications in clinical diagnosis of label-free fluorescent copper nanoclusters (CuNCs) are demonstrated. Double-strand DNA (dsDNA) can act as an effective template for the formation of CuNCs, which can be used to distinguish the deletion or duplication genotypes of Duchenne muscular dystrophy (DMD) due to different fluorescent intensities. After PCR, the DMD amplicons reacted with copper ion by reduction of ascorbic acid and generated fluorescence. The exons of the DMD gene were taken as the model analytes for genetic diagnosis. In this sensing system, the deletion type does not show fluorescence; on the other hand, the duplication type emits higher fluorescence than normal type. Parameters of this sensing system were optimized, including PCR conditions, levels of copper ion and ascorbate, and reaction time. The DMD-dominated exons 45, 46, and 47 were detected, and the method was applied to six samples of DMD patients. The results were consistent with those of the multiplex ligation-dependent probe amplification method. This strategy was feasible to detect all exons of this disease.
Original languageEnglish
Pages (from-to)6228-6232
Number of pages5
JournalAnalytical Chemistry
Volume87
Issue number12
DOIs
StatePublished - 16 Jun 2015

Keywords

  • POLYMERASE CHAIN-REACTION; DNA-TEMPLATED FORMATION; REAL-TIME PCR; DMD GENE; CAPILLARY-ELECTROPHORESIS; X-CHROMOSOME; SHORT ARM; PROBE; NANOPARTICLES; AMPLIFICATION

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