Screening for CHARGE syndrome mutations in the CHD7 gene using denaturing high-performance liquid chromatography

Michihiko Aramaki, Toru Udaka, Chiharu Torii, Hazuki Samejima, Rika Kosaki, Takao Takahashi, Kenjiro Kosaki

Research output: Contribution to journalArticle

9 Citations (Scopus)

Abstract

Mutations in the CHD7 (chromodomain helicase DNA binding protein 7) gene cause CHARGE syndrome. At present, however, genetic testing of the CHD7 gene is not commonly applied in clinical settings because the currently available assays are technically and financially demanding, mainly because of the size of the gene. In the present study, we optimized the highly sensitive and specific mutation scanning method automated de-naturing high-performance liquid chromatography (DHPLC) to analyze the entire coding region of CHD7. The coding region was amplified by 39 primer pairs, all of which have the same cycling conditions, aliquoted on a 96-well format polymerase chain reaction (PCR) plate. In this manner, all of the exons were amplified simultaneously using a single block in a thermal cycler. We then wrote a computer script to analyze each segment of the CHD7 gene by DHPLC in a serial manner using conditions that were optimized for each amplicon. The implementation of this screening method for CHD7 will help medical geneticists confirm their clinical impressions and provide accurate genetic counseling to the patients with CHARGE syndrome and their families.

Original languageEnglish
Pages (from-to)244-251
Number of pages8
JournalGenetic Testing
Volume10
Issue number4
DOIs
Publication statusPublished - 2006 Dec

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CHARGE Syndrome
DNA-Binding Proteins
High Pressure Liquid Chromatography
Mutation
Genes
Genetic Counseling
Genetic Testing
Exons
Hot Temperature
Polymerase Chain Reaction

ASJC Scopus subject areas

  • Genetics(clinical)

Cite this

Screening for CHARGE syndrome mutations in the CHD7 gene using denaturing high-performance liquid chromatography. / Aramaki, Michihiko; Udaka, Toru; Torii, Chiharu; Samejima, Hazuki; Kosaki, Rika; Takahashi, Takao; Kosaki, Kenjiro.

In: Genetic Testing, Vol. 10, No. 4, 12.2006, p. 244-251.

Research output: Contribution to journalArticle

Aramaki, Michihiko ; Udaka, Toru ; Torii, Chiharu ; Samejima, Hazuki ; Kosaki, Rika ; Takahashi, Takao ; Kosaki, Kenjiro. / Screening for CHARGE syndrome mutations in the CHD7 gene using denaturing high-performance liquid chromatography. In: Genetic Testing. 2006 ; Vol. 10, No. 4. pp. 244-251.
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