Pitfalls in the detection of gross gene rearrangements using MLPA in XXXX .

@entity1 @entity71071 @entity1112 @entity2314 @entity10153
UNASSIGNED: MLPA (Multiplex Ligation-dependent Probe Amplification) is a semiquantitative molecular technique developed to uncover gross gene rearrangements in several monogenic diseases, including the @entity1112 ( @entity2314 ) caused by mutations in the @entity10153 gene. Heterozygosity of the X chromosome in females makes it important to combine routine sequencing analysis with at least one allelic dosage assay (i.e. MLPA). We identified two new gross @entity10153 gene rearrangements, which were not properly detected by MLPA in male @entity1 with @entity2314 . In @entity1 1, routine sequencing did not amplify @entity10153 exon 7. MLPA failed to confirm such deletion. An alternative allele dosage, based on the Quantitative Fluorescent Multiplex-PCR (QFM-PCR), confirmed the mutation in the proband and excluded it in the mother, revealing that the mutation was de novo in the proband. @entity1 2 harboured a gross @entity10153 gene deletion encompassing almost the entire exon 5, the entire intron 5 and part of exon 6. MLPA confirmed the deletion of exon 5 but missed the partial deletion of exon 6. We characterised the breakpoint ( @entity71071 ) and carried out screening for possible heterozygosity among at risk female family members. MLPA can miss some gross @entity10153 gene rearrangements making the combination with other allele dosage assays mandatory to confirm or exclude @entity2314 at the molecular level.
