The role of polymorphic short tandem (CA)n repeat loci segregation analysis in the detection of Duchenne muscular dystrophy carriers and prenatal diagnosis.

Background: Duchenne and Becker muscular dystrophies (DMD/BMD) are X-linked diseases caused by mutations in the dystrophin gene at Xp21.2; they include gross deletions (60%), duplications (10%), and small mutations (30%). Since there is no cure or effective treatment for progressive muscular dystrophy, prevention of the disease is important and strongly depends on carrier-status information. Two-thirds of DMD/BMD cases are familial; thus, female relatives are candidates for carrier-risk assessment.

Aim: Segregation analysis of polymorphic short tandem (CA)n repeats [STR-(CA)n] was used to establish and compare the haplotypes of female relatives of patients with DMD/BMD with those of the patient in order to identify the mutant dystrophin gene and thus determine each female relative's carrier status.

Methods: 248 individuals from 52 families were studied through segregation of up to 11 STR-(CA)n loci. The assay was performed on leukocyte DNA by PCR amplification, polyacrylamide-gel electrophoresis and autoradiography. Haplotypes were established by determination of alleles on the autoradiography.

Results: 38 of 51 (75%) female relatives from familial cases were diagnosed as carriers or non-carriers with a 95-100% likelihood, and 18 out of 56 (32%) female relatives from sporadic cases could be excluded from the risk of being a DMD carrier with the same probability. In addition, STR studies detected gross deletions in 13 of the 52 (25%) families in both male and female individuals, four of which were de novo deletions. STR assays were also informative in families without an available DNA sample of an affected male and in two of seven symptomatic females. Determination of carrier status was particularly significant for prediction of DMD risk in prenatal analysis of five male chorionic villi. Other genetic events revealed by STR analysis were: (i) 11 recombinations identified in 6.6% of meiosis in the DMD families; (ii) germinal mosaicism detected in two female carriers; and (iii) changes in STR-(CA)n length during transmission from father to daughters, including three retractions and one elongation at an estimated rate of 0.004.

Conclusion: The STR assay is an excellent molecular tool for carrier-status identification and the detection of deletions and other genetic changes in families affected by DMD/BMD. Thus, it is useful in genetic counseling for the prevention of this disease.