Clin Exp Otorhinolaryngol.  2009 Mar;2(1):44-47. 10.3342/ceo.2009.2.1.44.

Construction of a DNA Chip for Screening of Genetic Hearing Loss

• Choi SY
• Kim YE
• Ahn DB
• Kim TH
• Choi JH
• Lee HR
• Hwang SJ
• Kim UK
• Lee SH

• KMID: 1466563
• DOI: http://doi.org/10.3342/ceo.2009.2.1.44

Abstract

OBJECTIVES
Hearing loss is the most common sensory disorder in humans and genetic causes are estimated to cause more than 50% of all incidents of congenital hearing loss. To develop an efficient method for a genetic diagnosis of hearing loss, we have developed and validated a genetic hearing loss DNA chip that allows the simultaneous analysis of 7 different mutations in the GJB2, SLC26A4, and the mtDNA 12S rRNA genes in Koreans.
METHODS
A genotyping microarray, based on the allele-specific primer extension (ASPE) method, was used and preliminary validation was examined from the five patients and five controls that were already known their genotypes by DNA sequencing analysis.
RESULTS
The cutoff Genotyping index (GI) of genotyping for each mutation was set up and validated to discriminate among the genotypes. The result of the DNA chip assay was identical to those of previous results.
CONCLUSION
We successfully designed the genetic hearing loss DNA chip for the first time in Korea and it would be useful for a clinical genetic diagnosis of hearing loss. Further consideration will be needed in order to examine the accuracy of this DNA chip with much larger patient sample numbers.

Keyword

Hearing loss; DNA chip; Gene

MeSH Terms

DNA
DNA, Mitochondrial
Genes, rRNA
Genotype
Hearing
Hearing Loss
Humans
Korea
Mass Screening
Oligonucleotide Array Sequence Analysis
RNA, Ribosomal
Sensation Disorders
Sequence Analysis, DNA
DNA
DNA, Mitochondrial
RNA, Ribosomal

Figure

• 

  Fig. 1 The principle of allele-specific primer extension (ASPE). ASPE is a solution based, sequence specific enzymatic reaction technology that can be used to assay multiple SNPs in a single tube. A primer anneals and is expended if there is a perfect match at the SNP site. The matched reaction is detected by fluorescence (Cy5) using scanner.

• 

  Fig. 2 Layout of the genetic hearing loss DNA chip. Oligonucleotides to wild-type sequences are indicated by a-N, and to mutant sequences by a-M.

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  Fig. 3 Examples of hybridization results. The genotype indexes calculated from the hybridization signals of mutant spots are also shown.

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