Genotyping microarray as a novel approach for the detection of ATP7B gene mutations in patients with Wilson disease
pmid: 18371106
Genotyping microarray as a novel approach for the detection of ATP7B gene mutations in patients with Wilson disease
Wilson disease (WD) is an autosomal recessive inherited disorder of copper metabolism that is caused by mutations in the ATP7B gene. To date, more than 300 mutations have been described in this gene. Molecular diagnostics of WD utilizes restriction enzyme digestion, multiplex ligation‐dependent probe amplification or a direct sequencing of the whole gene. To simplify and speed up the screening of ATP7B mutations, we have developed a genotyping microarray for the simultaneous detection of 87 mutations and 17 polymorphisms in the ATP7B gene based on the arrayed primer extension reaction. The patient’s DNA is amplified in four multiplex polymerase chain reactions, fragmented products are annealed to arrayed primers spotted on a chip, which enables DNA polymerase extension reactions with fluorescently labeled dideoxynucleotides. The Wilson microarray was validated by screening 97 previously genetically confirmed WD patients. In total, we detected 43 mutations and 15 polymorphisms that represent a majority of the common mutations occurring in the Czech and Slovak populations. All screened sequence variants were detected with 100% accuracy. The Wilson chip appears to be a rapid, sensitive and cost‐effective tool, representing the prototype of a disease chip that facilitates and speeds up the screening of potential WD patients.
- University Hospital Brno Czech Republic
- Masaryk University Czech Republic
- Asper Biotech (Estonia) Estonia
Adenosine Triphosphatases, Heterozygote, Genotype, Genetic Carrier Screening, DNA Mutational Analysis, Microarray Analysis, Hepatolenticular Degeneration, Copper-Transporting ATPases, Mutation, Humans, Point Mutation, Cation Transport Proteins
Adenosine Triphosphatases, Heterozygote, Genotype, Genetic Carrier Screening, DNA Mutational Analysis, Microarray Analysis, Hepatolenticular Degeneration, Copper-Transporting ATPases, Mutation, Humans, Point Mutation, Cation Transport Proteins
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