title: Mutation analysis of the pyruvate dehydrogenase E1 alpha gene in eight patients with a pyruvate dehydrogenase complex deficiency.
abstract: Most of the mutations causing deficiency of the pyruvate dehydrogenase (PDH) complex are in the X-linked E1 alpha gene. We have developed a rapid screening method for the detection of mutations in this gene using reverse transcription of total RNA, polymerase chain reaction amplification of the whole coding region of the gene and single-strand conformation polymorphism (SSCP) analysis. With this method, we studied eight patients with a PDH complex deficiency, using cultured fibroblasts. In all patients, aberrant SSCP patterns were found and, after sequencing of the corresponding fragments, we were able to identify six new mutations and two mutations already described previously. The mutations are point mutations leading to amino acid substitutions (5) and direct repeat insertions (3). The presence of the mutations was confirmed in genomic fibroblast DNA. The 4 female patients were shown to carry both a normal and a mutated E1 alpha gene.
authors.affiliation: Department of Medical Genetics, University Hospital, Brussels, Belgium.
authors.forename: W
authors.initials: W
authors.lastname: Lissens
authors.forename: L
authors.initials: L
authors.lastname: De Meirleir
authors.forename: S
authors.initials: S
authors.lastname: Seneca
authors.forename: C
authors.initials: C
authors.lastname: Benelli
authors.forename: C
authors.initials: C
authors.lastname: Marsac
authors.forename: B T
authors.initials: BT
authors.lastname: Poll-The
authors.forename: P
authors.initials: P
authors.lastname: Briones
authors.forename: W
authors.initials: W
authors.lastname: Ruitenbeek
authors.forename: O
authors.initials: O
authors.lastname: van Diggelen
authors.forename: D
authors.initials: D
authors.lastname: Chaigne
authors.forename: V
authors.initials: V
authors.lastname: Ramaekers
authors.forename: I
authors.initials: I
authors.lastname: Liebaers
mesh_terms: D001483:Base Sequence; D002478:Cells, Cultured; D002648:Child; D005260:Female; D005347:Fibroblasts; D006801:Humans; D007223:Infant; D008297:Male; D008969:Molecular Sequence Data; D009154:Mutation; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D018807:Polymorphism, Single-Stranded Conformational; D030481:Pyruvate Dehydrogenase (Lipoamide); D011768:Pyruvate Dehydrogenase Complex / Q000235:genetics*; D015325:Pyruvate Dehydrogenase Complex Deficiency Disease / Q000175:diagnosis / Q000235:genetics*; D012091:Repetitive Sequences, Nucleic Acid; D014158:Transcription, Genetic; D014960:X Chromosome
chemical_list: D011768:Pyruvate Dehydrogenase Complex; D030481:Pyruvate Dehydrogenase (Lipoamide); C065640:pyruvate dehydrogenase E1alpha subunit