DANALITICA LU UlOMifJlCUB www.-abBnjlitioB.il MULTIPLEX AMPLIFICATION INTRODUCTION Duchenne and Becker muscular dystrophies (DMD, BMD) are allelic neuromuscular disorders caused by mutations to the dystrophin gene atbXp21. Both involves a progressive and not-reversible degeneration of muscular fibers. Their incidences are respectively 1.5 per 10.000 and 1.5 per 100.000 live births. Because of the extremely large size of the dystrophin gene (more than 2.5Mb, 82 exons) several mutations are intragenic deletions (65%) or duplications (5-8%). DDK/G17 allows to detect the deletions in the exons 3, 4, 6, 8, 13, 17, 19, 43, 44, 45, 47, 48, 49, 50, 51, 52, 60 of the Duchenne and Becker muscular dystrophy gene, by amplification of this exons with three multiplex PCR. Traditionally, deletions and duplications of the dystrophin gene have been detected by Southern blot analysis using cDNA probes. These studies require big quantities and optimal samples of genomic DNA. The advent of PCR technology introduced the method of multiplex amplification of exonic sequences for detection of intragenic deletions. This method allows rapid (1-2 days) analysis on small quantities or suboptimal samples of DNA. This kit also allows to confirm the diagnosis in infected individuals and to do a prenatal diagnosis: it represents an important aid in genetic consulting for families. SHELF LIFE: 6 months. STARTING MATERIAL: peripheral whole blood. AMPLIFIED REGIONS: exons 3, 4, 6, 8, 13, 17, 19, 43, 44, 45, 47, 48, 49, 50, 51,52, 60. AMPLIFICATION: MULTIPLEX PCR. INTERNAL CONTROL: included. RESULT VISUALIZATION: agarose gel electrophoresis. Muntoni F. et al., Lancet Neurol. 2003 Dec;2(12):731-40. Navarro-Fdez Balbuena C. et al., Rev Neurol. 2003 Oct 1631 ;37(8):766-9 AB ANALITICA srl Via Svizzera 16 - 35127 PADOVA - ITALY Tel. +39 049 761698 - Fax +39 049 8709510 e-mail: [email protected] www.abanalitica.it