Combination of conventional multiplex PCR and quantitative real-time PCR detects large rearrangements in the dystrophin gene in 59% of Syrian DMD/BMD patients.
OBJECTIVE
Adaptation of a low-cost protocol to diagnose large rearrangements of the dystrophin gene in DMD/BMD Syrian patients and to establish the distribution of these mutations in the 2 hotspots.


METHODS
gDNA from 51 unrelated Syrian DMD/BMD male patients was isolated and analyzed by multiplex PCR of 25 hotspot exons in order to detect deletions. Patients who did not show any deletions were further analyzed by quantitative real-time PCR and the DeltaDeltaCt method in order to detect duplications in exons 4, 17, 47 and 52.


RESULTS
We found a deletion in 25 (49%) out of 51 patients studied. Quantitative real-time PCR revealed a duplication in 5 (9.8%) out of 51 patients. Combination of traditional multiplex PCR of hotspot exons with real-time PCR quantification of only exons 4, 17, 47 and 52 positively diagnosed 59% of Syrian DMD/BMD patients.


CONCLUSIONS
Our method may be useful as a cost-effective first-line test for the diagnosis of DMD/BMD patients before using exhaustive and expensive methods.
Madania
Ammar
A
Department of Radiation Medicine, Atomic Energy Commission, P.O. 6091, Damascus, Syria. scientific@aec.org.sy
Zarzour
Hana
H
Jarjour
Rami A
RA
Ghoury
Ifad
I
D000293:Adolescent; D000328:Adult; D002648:Child; D002675:Child, Preschool; D004252:DNA Mutational Analysis; D016189:Dystrophin / Q000235:genetics*; D006801:Humans; D007223:Infant; D008297:Male; D020388:Muscular Dystrophy, Duchenne / Q000175:diagnosis / Q000235:genetics*; D009154:Mutation / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods*; D013593:Syria
D016189:Dystrophin