title: Fluorescent multiplex PCR--fast method for autosomal dominant spinocerebellar ataxias screening.
abstract: Expansion of CAG trinucleotide repeats has been shown to cause a number of autosomal dominant spinocerebellar ataxias such as SCA1, SCA2, SCA3/MJD, SCA6 and SCA7. These disorders are characterized by a wide inter- and intrafamiliar variation in clinical features. The same mutation can result in different phenotypes and the very similar phenotypes can be caused by different mutations. Therefore it is necessary to investigate more SCA genes (according to prevalence) to identify the causal elongation. We developed a fast and efficient screening method based on touchdown multiplex PCR with fluorescent labelled primers for the most common types of SCAs (SCA 1, 2, 3 and 7). It has been reliable in 113 probands tested. Fragment analysis was performed by using 6% denaturing polyacrylamide gel and employing the automated DNA sequencer. This method considerably shortens the process of molecular genetic screening of SCAs and might be used as a tip for designing other SCA screening sets.
authors.affiliation: Neurogenetic Center, Institute of Biology and Medical Genetics, Department of Child Neurology, 2nd Medical Faculty of Charles University, Faculty Hospital Motol, Prague 150 06, Czech Republic. pbauer@brain.riken.go.jp
authors.forename: P O
authors.initials: PO
authors.lastname: Bauer
authors.forename: S E
authors.initials: SE
authors.lastname: Kotliarova
authors.forename: V
authors.initials: V
authors.lastname: Matoska
authors.forename: Z
authors.initials: Z
authors.lastname: Musova
authors.forename: P
authors.initials: P
authors.lastname: Hedvicakova
authors.forename: A
authors.initials: A
authors.lastname: Boday
authors.forename: A
authors.initials: A
authors.lastname: Tomek
authors.forename: N
authors.initials: N
authors.lastname: Nukina
authors.forename: P
authors.initials: P
authors.lastname: Goetz
mesh_terms: D005820:Genetic Testing / Q000379:methods; D006801:Humans; D016133:Polymerase Chain Reaction / Q000379:methods; D020754:Spinocerebellar Ataxias / Q000235:genetics*; D019680:Trinucleotide Repeat Expansion / Q000235:genetics*