Document Type : Original Article
PASTEUR INSTITUTE, BIOTECHNOLOGY RESEARCH CENTER, TEHRAN, IRAN
Introduction: Duchene/ Becker (DMD/BMD) muscular dystrophy is the most frequent neuromuscular disease in children which is inherited as an X-Linked recessive trait. The disease is caused by partial deletion in dystrophin gene. We developed a rapid and robust method for direct identification of female carriers of deletions and duplications in the dystrophin gene, in order to prevent the affected newborn males to be born.
Materials and methods: Qantitative real-time PCR assay for deletion/duplication within two major hot spots in dystrophin gene (exon 6 and 47) was developed and an X-linked gene was considered as a normalizer (i.e., a fragment of human coagulation factor IX gene).The quantitative Real-time PCR test was conducted using comparative threshold method (ΔΔCT).
Results: The results showed 2-ΔΔCT of 1.09±0.21 for normal individuals and 0.51±0.1 for carrier samples. Carrier status was accurately attributed in 100% of cases.
Conclusion: This method is simple, rapid, reliable and cost-effective. It may be applied for direct determination of deletion/duplication in potential DMD/BMD carriers and may easily be adapted for other genetic conditions involving deletions and duplications.