DYSTROPHIN GENE ANALYSIS ON 130 PATIENTS WITH DUCHENNE MUSCULAR-DYSTROPHY WITH A SPECIAL REFERENCE TO MUSCLE MESSENGER-RNA ANALYSIS

On dystrophin gene analysis by multiplex polymerase chain reaction (PC R), 76 of 130 (58.5%) Japanese patients with Duchenne muscular dystrop hy had a deletion or duplication in genomic DNA. Of the remaining 54 p atients who had no identifiable gene mutations, muscle biopsy tissue w as available in 16 for RNA extraction. The full length of the coding r egions of dystrophin cDNA was amplified in IO fragments by reverse tra nscription nested PCR (RT-PCR). Five of 16 patients (31%) had dystroph in cDNA of abnormal size. One patient had a deletion, and two duplicat ions that were not covered by multiplex PCR; one an exon-skipping of e xon 51 caused by a 5' consensus splice site mutation of intron 51, and one 172 bp or 202 bp insertion in the cDNA between exon 25 and 26. Ne sted RT-PCR from the total RNA extracted from muscle biopsy was useful for screening patients who had no identifiable gene abnormality by mu ltiplex PCR. (C) 1998 Elsevier Science B.V.