title: Evaluation of IP-RP-HPLC for length determination of the trinucleotide repeat fragments in Huntington's disease.
abstract: Expansion of an unstable trinucleotide (CAG)(n) repeat region within exon 1 of the gene IT15 causes autosomal, dominantly inherited Huntington's disease (HD). The number of CAG-repeats varies from 6 to 35 in normal individuals, whereas in affected patients the expanded allele contains 40 or more CAG-repeats. Thus, exact determination of both alleles of the gene (normal and expanded) on the molecular level is of great importance for clinical diagnosis and prognosis regarding the course of the disease. In our study, we optimized and evaluated a highly sensitive, automated, and economical molecular method for length characterization of the trinucleotide fragment expansion such as (CAG)(n) repeat region based on ion-pair reversed-phase high-performance liquid chromatography (IP-RP-HPLC). We found that IP-RP-HPLC can be used for exact fragment length measuring between 60-280 bp as a sensitive and advantageous alternative method to conventional techniques.
authors.affiliation: Institute of Human Genetics, University of Leipzig, Philipp-Rosenthal-Str. 55, 04103 Leipzig, Germany.
authors.forename: Marzena
authors.initials: M
authors.lastname: Skrzypczak
authors.forename: Wolfram
authors.initials: W
authors.lastname: Heinritz
authors.forename: Anna M
authors.initials: AM
authors.lastname: Schulz
authors.forename: Marek
authors.initials: M
authors.lastname: Mierzejewski
authors.forename: Ursula G
authors.initials: UG
authors.lastname: Froster
mesh_terms: D000483:Alleles; D002851:Chromatography, High Pressure Liquid / Q000191:economics / Q000379:methods*; D056148:Chromatography, Reverse-Phase / Q000191:economics / Q000379:methods*; D004247:DNA / Q000032:analysis* / Q000235:genetics; D006801:Humans; D006816:Huntington Disease / Q000175:diagnosis* / Q000235:genetics*; D012680:Sensitivity and Specificity; D019680:Trinucleotide Repeat Expansion
chemical_list: D004247:DNA