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DNA analysis of Brazilian Duchenne Muscular Dystrophy families using (CA)n microsatellite markers

Daisy Neves Falcão-Conceição^I; Alexander L.J. Kneppers^II; E. Bakker^II

^IDepartamento de Genética, Centro de Ciências da Saúde, Bloco A-I, Biologia, Universidade Federal do Rio de Janeiro, 21941-900 Rio de Janeiro, RJ, Brasil. Send correspondence to D.N.F.-C.
^IIDepartament of Human Genetics, University of Leiden, Leiden, The Netherlands

ABSTRACT

We investigated nine Brazilian Duchenne Muscular Dystrophy (DMD) families with patients without a deletion or duplication, by haplotype analysis using microsatellite markers of the (CA)n type. Our objective was to detect carriers of the anomalous dystrophin gene among women at risk. We assumed that the locus Xp21 is involved in the MD condition in all the families. Looking for informativeness, we used two (CA)n microsatellite markers of the 5' end of the dystrophin gene, i.e. 5'DYSI and 5'DYSIII (Feener et al., Am. J. Hum. Genet. 48: 621-627, 1991) and, in view of the high intragenic recombination rate, we also used a microsatellite marker of the 49 intron, i.e., STR 49 (Clemens et al., Am. J. Hum. Genet. 49: 951-960, 1991). In only one of the nine families the information was insufficient to indict the X-chromosome at risk. There was absolute coherence between increased creatine-kinase (CK) levels of some obligatory carrier women and their genetic condition as evaluated by haplotype analysis. In three of seven families with isolated cases it was possible to define new mutations in which the X-chromosome was originated from the maternal grandfather. It was possible to detect a crossing over at the 5' end-intron 49 interval in one of the nine mothers of patients.
We conclude that (CA)n loci are excellent markers in face of their high informativeness. Sensitivity and rapidity provided by the polymerase chain reaction (PCR) assay make them unique when compared to other markers assayed by standard Southern blotting and hybridization. For detection of carriers of the anomalous dystrophin gene, it is absolutely necessary to use markers both from 5' or 3' end of the gene and from the central/distal (3') regions in face of the 12% intragenic recombination rate (Abbs et al., Genomics 7: 602-606, 1990).

Keywords: DNA; duchenne; muscular dystrophy; microsatellite markers.

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