Multiplex Ligation-dependent Probe Amplification Analysis Subsequent to Direct DNA Full Sequencing for Identifying ATP7B Mutations and Phenotype Correlations in Children with Wilson Disease

• Published in: Journal of Korean medical science Vol. 33; no. 26; p. e177
• Main Authors: Shim, Jung Ok, Yang, Hye Ran, Moon, Jin Soo, Chang, Ju Young, Ko, Jae Sung, Park, Sung Sup, Seo, Jeong Kee
• Format: Journal Article
• Language: English
• Published: Korea (South) The Korean Academy of Medical Sciences 25.06.2018; 대한의학회
• Subjects: Child; DNA; DNA Mutational Analysis; Exons; Hepatolenticular Degeneration - genetics; Humans; Multiplex Polymerase Chain Reaction; Mutation; Original; Phenotype; 의학일반; Mutation; Sequence Analysis; Multiplex Ligation-dependent Probe Amplification; Child; Wilson Disease
• DOI: 10.3346/JKMS.2018.33.E177
• ISSN: 1011-8934

Mutations in cause Wilson disease (WD). However, direct DNA full sequencing cannot detect all mutations in patients with WD. Multiplex ligation-dependent probe amplification (MLPA) analysis is reportedly useful in increasing the diagnostic yield in other genetic disorders with large deletions or insertions. The aim of this study was to evaluate whether the detection rate of mutations can be increased by using MLPA. We enrolled 114 children with WD from 104 unrelated families based on biochemical tests and direct DNA full sequencing. The patients with one or zero mutant allele were investigated using MLPA. We analyzed phenotypic correlations. Total allele frequency by full sequencing was 87.5%. Full sequencing revealed two mutant alleles in 80 of 104 unrelated children. One mutant allele was detected in 22 children, and no mutations were found in two children. Novel mutations including small deletions with frameshift mutations were identified by DNA sequencing. MLPA revealed no gross deletion or duplication in 24 children with one or zero mutant alleles. The number of detected mutations was not associated with hepatic manifestation, age of onset, Kayser-Fleischer ring, ceruloplasmin, and urinary Cu concentrations. MLPA showed a limited role to increase the mutation detection rate in children who do not receive a definite genetic diagnosis of WD through DNA full sequencing. This finding suggests that large deletions or duplications might be extremely rare in WD. Further development is needed to improve the genetic diagnosis of WD.