Efficient Detection of Mutations in Wilson Disease by Manifold Sequencing |
| |
Authors: | Erik Waldenströ m,Arild Lagerkvist,Thé rè se Dahlman,Kerstin Westermark,Ulf Landegren |
| |
Affiliation: | aThe Beijer Laboratory, Department of Medical Genetics, Uppsala Biomedical Center, S-751 23, Uppsala, Sweden;bDepartment of Internal Medicine, University Hospital, S-751 85, Uppsala, Sweden |
| |
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. |
| |
Keywords: | |
本文献已被 ScienceDirect 等数据库收录! |
|