Mutation analysis of the pyruvate dehydrogenase E1α gene in eight patients with a pyruvate dehydrogenase complex deficiency

✍ Scribed by Willy Lissens; Linda De Meirleir; Sara Seneca; Chantal Benelli; Cécile Marsac; Bwee Tien Poll-The; Paz Briones; Wim Ruitenbeek; Otto van Diggelen; Denis Chaigne; Vincent Ramaekers; Ingeborg Liebaers

• Publisher: John Wiley and Sons
• Year: 1996
• Tongue: English
• Weight: 598 KB
• Volume: 7
• Category: Article
• ISSN: 1059-7794
• DOI: 10.1002/(sici)1098-1004(1996)7:1<46::aid-humu6>3.0.co;2-n

✦ Synopsis

Most of the mutations causing deficiency of the pyruvate dehydrogenase (PDH) complex are in the X-linked E,a 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). T h e presence of the mutations was confirmed in genomic fibroblast DNA. T h e 4 female patients were shown to carry both a normal and a mutated E , a gene. o 19% WiIey-Liss, Inc.