Résumé :
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Communication n° 427 Introduction : Duchenne and Becker muscular dystrophy (DMD/BMD) are common X-chromosomal recessive disorders caused by mutations in the dystrophin gene. The majority (2/3) of recognized mutations are copy number changes in one of the 79 individual exons. Objectives : Here we report the results of dystrophin gene analysis of Hungarian DMD/BMD patients performed by different molecular diagnostic techniques. Methods : Detection of deletion patterns in 103 male patients was first performed by multiplex PCR technique that enables simultaneous screening of the promoter Pm and 17 exons of the dystrophin gene. In 59 cases deletions were detected in the most commonly affected exons. Identification of female carriers was carried out by radioactive Southern blot hybridization using several cDNA probes. Out of the 32 female family members, where deletions were detected in the index patients, 17 proved to be carriers of deletions of the DMD/BMD gene. Recently, we introduced the newly developed multiplex ligation-dependent probe amplification (MLPA) assay. Results : Until now, 10 male patients have been analyzed and the method developed for the agarose gel detection confirmed all previously recognized mutations and identified additional ones in four patients, thus determining the exact deletion borders. Moreover, in one patient without any previous deletion history, rare exon deletions were detected. To improve the MLPA assay, we also introduced quantitative fluorescent detection by a capillary electrophoresis system that will allow us to detect additional duplications and to recognize female carriers easier than by the previous cDNA assay. These results will be discussed on the poster. Conclusions : According to our preliminary data, the use of MLPA analysis will facilitate and improve the diagnosis of DMD/BMD patients in Hungary as the mutation pick-up rate will be increased significantly.
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