title: Efficient detection of mutations in Wilson disease by manifold sequencing.
abstract: We have applied a solid support for parallel handling and direct loading of sequencing reactions--manifold sequencing--to analyze the coding sequence for the deficient copper transporting P-type ATPase in 24 families with Wilson disease. At least 100 different amplification reactions could be handled in parallel, with a minimal turnaround time of 12 h from isolated genomic DNA to identification of the mutations. Sixteen different mutations were found, accounting for 92% of the mutant genes. Ten of these mutations have not been previously described. Eleven were observed only in single families. Mutation His1069Gln, previously identified as the most prevalent mutation in Northern Europe, was found in one-third of the Northern European chromosomes in our material. Four patients were homozygous for this mutation, and three were homozygous for Thr977Met. The method allowed us to establish the diagnosis of Wilson disease in 24 h in a patient with acute hepatic failure.
authors.affiliation: Beijer Laboratory, Department of Medical Genetics, Uppsala Biomedical Center, Sweden. erik.waldenstrom@medgen.uu.se
authors.forename: E
authors.initials: E
authors.lastname: Waldenström
authors.forename: A
authors.initials: A
authors.lastname: Lagerkvist
authors.forename: T
authors.initials: T
authors.lastname: Dahlman
authors.forename: K
authors.initials: K
authors.lastname: Westermark
authors.forename: U
authors.initials: U
authors.lastname: Landegren
mesh_terms: D000251:Adenosine Triphosphatases / Q000235:genetics*; D000293:Adolescent; D000328:Adult; D017668:Age of Onset; D002352:Carrier Proteins / Q000235:genetics*; D027682:Cation Transport Proteins; D002648:Child; D000073840:Copper-Transporting ATPases; D004252:DNA Mutational Analysis / Q000295:instrumentation / Q000379:methods*; D004262:DNA Restriction Enzymes; D005091:Exons / Q000235:genetics; D005260:Female; D006527:Hepatolenticular Degeneration / Q000150:complications / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*; D006801:Humans; D017093:Liver Failure / Q000209:etiology; D008297:Male; D008875:Middle Aged; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D017422:Sequence Analysis, DNA / Q000295:instrumentation / Q000379:methods*
chemical_list: D002352:Carrier Proteins; D027682:Cation Transport Proteins; D004262:DNA Restriction Enzymes; D000251:Adenosine Triphosphatases; D000073840:Copper-Transporting ATPases