Validation and comparison of two quantitative real-time PCR assays for direct detection of DMD/BMD carriers

Objectives: To develop a robust and reliable assay for direct identification of female carriers of deletions in the dystrophin gene. Design and methods: We compared two quantitative real-time PCR approaches for the detection of the deletions of exons 4, 17, 47, and 50 in DMD/BMD carriers. One hundred and ten individuals from 26 unrelated families, including 8 large pedigrees characterized by having at least two DMD affected males, were studied. Carrier status of the subjects was also evaluated by MLPA. Results: The results showed the gene dosage ratio of 0.99±0.14 and 1.09±0.19 for normal individuals and 0.48±0.06 and 0.50±0.10 for carriers in SYBR green and TaqMan probe assays, respectively. Carrier status was accurately attributed in 100% of cases and confirmed by MLPA. Conclusion: Quantitative real-time PCR can be used as a direct method for carrier detection in female relatives of DMD patients with known deletions. The results are comparable to the MLPA data.