title: Comparison of different techniques for detecting 17p12 duplication in CMT1A.
abstract: Charcot-Marie-Tooth type 1A is caused by a 1.5Mb DNA duplication in the 17p12 chromosomal region encompassing the peripheral myelin protein 22 gene. In the present study, we compared the Real-Time PCR with the other methods currently used for the diagnosis of Charcot-Marie-Tooth. By using a combination of junction fragment PCR, analysis of microsatellite markers, and pulsed field gel electrophoresis, we identified 76 unrelated patients with 17p12 duplication. In these patients, junction fragment PCR detected 63% of cases of duplication, the microsatellite markers method revealed 74%, while the combined use of microsatellite markers and junction fragment PCR revealed 91% of cases of Charcot-Marie-Tooth type 1A. Pulsed field gel electrophoresis detected 100% of the cases with duplication, even in presence of atypical 17p12 duplication. Real-Time PCR detected 100% of the cases with Charcot-Marie-Tooth type 1A and was comparable to pulsed field gel electrophoresis. However, in contrast to pulsed field gel electrophoresis, Real-Time PCR does not need fresh blood, minimizes diagnosis time and cost, and thus can be easily used for the molecular diagnosis of Charcot-Marie-Tooth type 1A.
authors.affiliation: Institute of Neurological Sciences, National Research Council, Piano Lago di Mangone, Cosenza, Italy.
authors.forename: Alessandra
authors.initials: A
authors.lastname: Patitucci
authors.forename: Maria
authors.initials: M
authors.lastname: Muglia
authors.forename: Angela
authors.initials: A
authors.lastname: Magariello
authors.forename: Anna Lia
authors.initials: AL
authors.lastname: Gabriele
authors.forename: Giuseppina
authors.initials: G
authors.lastname: Peluso
authors.forename: Teresa
authors.initials: T
authors.lastname: Sprovieri
authors.forename: Francesca Luisa
authors.initials: FL
authors.lastname: Conforti
authors.forename: Rosalucia
authors.initials: R
authors.lastname: Mazzei
authors.forename: Carmine
authors.initials: C
authors.lastname: Ungaro
authors.forename: Francesca
authors.initials: F
authors.lastname: Condino
authors.forename: Paola
authors.initials: P
authors.lastname: Valentino
authors.forename: Franco
authors.initials: F
authors.lastname: Bono
authors.forename: Carmelo
authors.initials: C
authors.lastname: Rodolico
authors.forename: Anna
authors.initials: A
authors.lastname: Mazzeo
authors.forename: Antonio
authors.initials: A
authors.lastname: Toscano
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Vita
authors.forename: Aldo
authors.initials: A
authors.lastname: Quattrone
mesh_terms: D015139:Blotting, Southern; D002607:Charcot-Marie-Tooth Disease / Q000175:diagnosis / Q000235:genetics*; D002886:Chromosomes, Human, Pair 17; D004252:DNA Mutational Analysis; D016521:Electrophoresis, Gel, Pulsed-Field / Q000379:methods; D020440:Gene Duplication; D006801:Humans; D018895:Microsatellite Repeats / Q000502:physiology; D009693:Nucleic Acid Hybridization / Q000379:methods; D012333:RNA, Messenger / Q000096:biosynthesis; D015203:Reproducibility of Results; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D012680:Sensitivity and Specificity; D018709:Statistics, Nonparametric
chemical_list: D012333:RNA, Messenger