Comparison of different techniques for detecting 17p12 duplication in CMT1A

• Published in: Neuromuscular disorders : NMD Vol. 15; no. 7; pp. 488 - 492
• Main Authors: Patitucci, Alessandra, Muglia, Maria, Magariello, Angela, Gabriele, Anna Lia, Peluso, Giuseppina, Sprovieri, Teresa, Conforti, Francesca Luisa, Mazzei, Rosalucia, Ungaro, Carmine, Condino, Francesca, Valentino, Paola, Bono, Franco, Rodolico, Carmelo, Mazzeo, Anna, Toscano, Antonio, Vita, Giuseppe, Quattrone, Aldo
• Format: Journal Article
• Language: English
• Published: England Elsevier B.V 01-07-2005
• Subjects: Blotting, Southern; Charcot-Marie-Tooth Disease - diagnosis; Charcot-Marie-Tooth Disease - genetics; Chromosomes, Human, Pair 17; CMT1A; DNA Mutational Analysis; Electrophoresis, Gel, Pulsed-Field - methods; Gene Duplication; Humans; Microsatellite Repeats - physiology; Nucleic Acid Hybridization - methods; PFGE; Real-Time PCR; Reproducibility of Results; Reverse Transcriptase Polymerase Chain Reaction - methods; RNA, Messenger - biosynthesis; Sensitivity and Specificity; Statistics, Nonparametric; PFGE; Real-Time PCR; CMT1A
• ISSN: 0960-8966; 1873-2364
• DOI: 10.1016/j.nmd.2005.04.006

Charcot–Marie-Tooth type 1A is caused by a 1.5
Mb 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.