Open AccessQuantitative analysis of the dystrophin gene by real-time PCR
Author(s) -
Nela Maksimović,
Ana Andjelković,
Vedrana Milić-Rašić,
Vidosava Rakočević-Stojanović,
Biljana Kastratović-Kotlica,
Slavko Brankovic,
Tatjana Damnjanović,
Biljana Jekić,
Vera Bunjevački,
Ljiljana Luković,
Dijana Perović,
Suzana Cvjetićanin,
Ivaovaković
Publication year - 2012
Publication title -
archives of biological sciences
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.217
H-Index - 25
eISSN - 1821-4339
pISSN - 0354-4664
DOI - 10.2298/abs1202787m
Subject(s) - duchenne muscular dystrophy , dystrophin , exon , gene , muscular dystrophy , real time polymerase chain reaction , biology , microbiology and biotechnology , genetics
Duchenne and Becker muscular dystrophy (DMD/BMD) are severe X-linked neuromuscular disorders caused by mutations in the dystrophin gene. Our aim was to optimize a quantitative real-time PCR method based on SYBR® Green I chemistry for routine diagnostics of DMD/BMD deletion carriers. Twenty female relatives of DMD/BMD patients with previously detected partial gene deletions were studied. The relative quantity of the target exons was calculated by a comparative threshold cycle method (ΔΔCt). The carrier status of all subjects was successfully determined. The gene dosage ratio for non-carriers was 1.07±0.20, and for carriers 0.56±0.11. This assay proved to be simple, rapid, reliable and cost-effective