Heiko Witt, Olfert Landt
Rapid Detection of the Wilson's Disease H1069Q Mutation by Melting Curve Analysis with the LightCycler
Wilson's disease is an inherited autosomal recessive
disorder of copper transport characterized by progressive copper accumulation in
the liver and the central nervous system. The disease is caused by mutations in
the ATP7B gene. Although many different mutations in this gene were
described, a substitution of a histidine by a glutamine residue at codon 1069
(H1069Q) accounts for approximately 30–60% of all mutations in Caucasian
patients. We describe a DNA-based method using fluorescence resonance energy
transfer probes on the LightCycler for rapid determination of the common H1069Q
mutation in the ATP7B gene. We screened 53 patients with Wilson's
disease for the H1069Q mutation by the melting curve analysis. The reliability
and discriminating power of this technique were documented by comparing results
of the LightCycler assay with direct DNA sequencing. The protocol allows
genotyping of 30 samples in less than 1 hour without a need for restriction
enzyme digestion or gel electrophoresis.
Clinical Chemical Laboratory Medicine, Walter de Gruyter
Print ISSN: 1434-6621
Volume: 39, 10/2001
Pages: 953 - 955
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