ABSTRACT
Carrier status determination for Duchenne and Becker muscular dystrophies (D/BMD), disorders caused by mutations in the dystrophin gene at Xp21, is complicated by a number of factors. These include a high mutation rate and a 5-10% recombination frequency across the dystrophin gene. For these reasons, linkage analysis frequently gives an inconclusive result, and a direct mutation detection method for females at risk is desirable. Because 65% of the mutations that cause D/BMD are deletions of one or more exons of the dystrophin gene, diagnosis in most affected males is relatively easy using multiplex polymerase chain reaction (PCR) analysis. However, deletion analysis in females is more difficult because of the interference of the normal X chromosome in the deletion assay. We have developed a quantitative PCR-based analysis designated computer-assisted laser densitometry (CALD), which uses the automated fluorescent fragment analysis application of the Applied Biosystems (Foster City, California) automated sequencer. This method has proved to be 100% accurate in retrospective blind studies analysing a total of 351 samples. Subsequent analysis of more than 800 women from more than 400 D/BMD families has shown that a highly accurate carrier risk can be given in more than 90% of cases.