title: Use of denaturing HPLC to provide efficient detection of mutations causing guanidinoacetate methyltransferase deficiency.
abstract: Guanidinoacetate methyltransferase (GAMT) deficiency is an autosomal recessive error of creatine synthesis characterized by cerebral creatine deficiency, accumulation of guanidinoacetate, mental retardation, epilepsy, and extrapyramidal symptoms. To date, 14 mutations of the GAMT gene in 27 patients have been reported. Mutation analysis was done using direct sequencing of PCR products and denaturing gradient gel electrophoresis in combination with direct sequencing. In contrast, we evaluated the efficiency of a newly developed DHPLC method to detect mutations in the GAMT gene by analysing DNA from 14 GAMT patients with known mutations. PCR amplification of both patient and control DNA was followed by formation of homoduplices and heteroduplices, and their detection by DHPLC. DHPLC identified all mutations tested and is the preferred choice of analytical method.
authors.affiliation: Biochemical Genetics and Newborn Screening Laboratories, University Children's Hospital, Vienna, Austria.
authors.forename: C B
authors.initials: CB
authors.lastname: Item
authors.forename: S
authors.initials: S
authors.lastname: Stöckler-Ipsiroglu
authors.forename: C
authors.initials: C
authors.lastname: Willheim
authors.forename: A
authors.initials: A
authors.lastname: Mühl
authors.forename: O A
authors.initials: OA
authors.lastname: Bodamer
mesh_terms: D001483:Base Sequence; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D017931:DNA Primers; D050937:Guanidinoacetate N-Methyltransferase / Q000172:deficiency / Q000235:genetics*; D009154:Mutation; D009691:Nucleic Acid Denaturation; D009692:Nucleic Acid Heteroduplexes; D016133:Polymerase Chain Reaction
chemical_list: D017931:DNA Primers; D009692:Nucleic Acid Heteroduplexes; D050937:Guanidinoacetate N-Methyltransferase