BASE SUBSTITUTIONS IN THE HUMAN DYSTROPHIN GENE - DETECTION BY USING THE SINGLE-STRAND CONFORMATION POLYMORPHISM (SSCP) TECHNIQUE

We have established the experimental conditions to screen twenty regio ns of the dystrophin gene using the method of single-strand conformati onal polymorphism (SSCP) analysis. The aim of this study was to identi fy point mutations in patients with Duchenne or Becker muscular dystro phy (DMD or BMD) who have no gross DNA rearrangements detectable by So uthern blot analysis or multiplex exon amplification. The investigatio n of thirteen patients using this procedure resulted in the detection of seven sequence polymorphisms (four identified in this study) that w ill be useful allelic markers in familial DNA analysis. Three rare seq uence variants could be found (two of them being novel variants) but w e were unable to demonstrate mutations that could be clearly sufficien t to be responsible for the phenotype. This analysis confirmed the eff iciency of the SSCP technique for the detection of nucleotide substitu tions. Application of this approach to mutation or polymorphism detect ion to other exons of the gene will improve carrier and prenatal diagn osis. (C) 1993 Wiley-Liss, Inc.