Identification of deletions and duplications of the DMD gene in affected males and carrier females by multiple ligation probe amplification (MLPA).
Duchenne muscular dystrophy (DMD) and Becker muscular dystrophy (BMD) are caused in the majority of cases by deletions of the DMD gene and are readily detectable in affected males by multiplex polymerase chain reaction (PCR). However, different approaches must be used for the identification of female carriers, in which deletions are not detectable by PCR, because of the presence of a normal X chromosome. In this study, we used the multiple ligation probe amplification (MLPA) tool for the identification of female carriers of DMD deletions or duplications in 12 families with a single affected male, 10 of which were previously diagnosed as carriers of a DMD rearrangement, and the remaining two as having an unknown disease-causing mutation. In all the investigated affected males, MLPA analysis confirmed the presence of a DMD rearrangement, and in six of them allowed the refinement of the breakpoints. In 12 female relatives of the affected patients, MLPA analysis showed a DMD deletion or duplication, confirming their carrier status. Two of these were the mother and the sister of a patient whose disease-causing mutation was not known. MLPA analysis thus proved to be an useful tool for the analysis of both affected males and females carriers of DMD rearrangements in cases in which the disease-causing mutation in the affected male was not known, providing useful information for the genetic counselling of the family.
Gatta
Valentina
V
Department of Biomedical Sciences, G. D'Annunzio University Foundation, Chieti, Italy.
Scarciolla
Oronzo
O
Gaspari
Anna Rita
AR
Palka
Chiara
C
De Angelis
Maria Vittoria
MV
Di Muzio
Antonio
A
Guanciali-Franchi
Paolo
P
Calabrese
Giuseppe
G
Uncini
Antonino
A
Stuppia
Liborio
L
D041321:Chromosomes, Human, X; D004252:DNA Mutational Analysis / Q000379:methods*; D015342:DNA Probes; D016189:Dystrophin / Q000235:genetics*; D005260:Female; D017353:Gene Deletion; D020440:Gene Duplication; D005817:Genetic Counseling; D006579:Heterozygote; D006801:Humans; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis* / Q000235:genetics*; D021141:Nucleic Acid Amplification Techniques; D010375:Pedigree; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity
C484258:DMD protein, human; D015342:DNA Probes; D016189:Dystrophin