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Screening for deletions in duchenne and becker muscular dystrophu patients using the genomic probe P20 and the cDNA XJ10 (1-2b), 7b-8 and 44-1 (8) probes

Daisy Neves Falcão-Conceição; Márcia M. Gonçalves-Pimentel; Cláudia Pimenta Moreira

Departamento de Genética, Instituto de Biologia, Universidade Federal do Rio de Janeiro, Caixa Postal 68011, Ilha do Fundão, 21944 Rio de Janeiro, RJ, Brasil

ABSTRACT

A study was carried out on 20 patients with Duchenne muscular dystrophy (DMD) and on 4 patients with Becker muscular dystrophy (BMD) with the objective of detecting deletions and duplications on the 5' end of the dystropin gene with the eDNA XJ10 (1-2b) probe and in the central region with the cDNA 7b-8 and 44-1 (8) probes.
All seven deletions and one duplication in the DMD patients were detected in the central region of the gene with the 44-1 (8) probe, while The two BMD deletions were better detected with the 7b-8 probe. The genomic probe P20 permitted the proximal delimitation of the extent of these deletions. The XJ10 (1-2b) probe detected one deletion and one duplication in DMD patients. Thus, 12 genomic alterations were detected by Southern blotting in a total of 24 patients (50%). In the present study, the polymerise chain reaction (PCR) analysis was of great help for confirming the presence or absence of low molecular weight fragments which were lost in some runs.
It was confirmed that the 8 (44-1) probe is ideal for the detection of DMD deletions and the 7b-8 probe ideal for the detection of both DMD and BMD deletions, since most of the BMD deletions involve exons 45 and 46, with a break point at P20, a region contained in a giant intron (44-45) of 170 kb. A possible hotspot (XJ) was observed in the two genomic alterations detected with the XJ10 probe (intron 7-8).

Keywords: Duchenne; Becker; muscular dystrophy.

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