Combination of conventional multiplex PCR and quantitative real-time PCR detects large rearrangements in the dystrophin gene in 59% of Syrian DMD/BMD patients

Objectives: 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. Design and 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 Delta Delta Ct 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. Conclusion: 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. (c) 2010 The Canadian Society of Clinical Chemists. Published by Elsevier Inc. All rights reserved.