Automated mutation screening using dideoxy fingerprinting and capillary array electrophoresis.
Hum Mutat
; 18(5): 451-7, 2001 Nov.
Article
em En
| MEDLINE
| ID: mdl-11668638
The rapid progress in the isolation of genes associated with human disease has resulted in an increasing demand for mutation screening methods. The molecular diagnosis of the long QT syndrome (LQTS), a cardiac disorder characterized by prolongation of the QT(c) interval in the ECG, syncopes, and sudden death, requires mutation screening of all exons in at least five genes, encoding cardiac Na(+) and K(+) channel subunits. A method for automated dideoxy fingerprinting (ddF) using capillary array electrophoresis (CAE) was developed and the efficiency of the method was tested by analyzing 24 DNA samples with mutations in one of the genes KCNQ1 and KCNH2, which are involved in 50% of LQTS cases. One of these mutations, 362insQK in KCNQ1, is novel. The sensitivity was 100% using a single electrophoresis temperature of 18 degrees C or 25 degrees C. However, analysis of the samples in both the sense and anti-sense direction were required for high sensitivity. Analysis in a single direction resulted in a decrease of the sensitivity to 74% and 70%, respectively. The throughput of the ddF method, if performed with a 16 capillary CAE instrument, is 288 samples per seven hr if each sample is analyzed on both strands.
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Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Didesoxinucleosídeos
/
Canais de Potássio
/
Transativadores
/
Testes Genéticos
/
Impressões Digitais de DNA
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Eletroforese Capilar
/
Canais de Potássio de Abertura Dependente da Tensão da Membrana
/
Proteínas de Transporte de Cátions
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Proteínas de Ligação a DNA
/
Mutação
Tipo de estudo:
Diagnostic_studies
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Prognostic_studies
/
Screening_studies
Limite:
Humans
Idioma:
En
Ano de publicação:
2001
Tipo de documento:
Article