A SENSITIVE MUTATION SCREENING STRATEGY FOR FABRY DISEASE - DETECTIONOF 9 MUTATIONS IN THE ALPHA-GALACTOSIDASE-A GENE

Fabry disease is an X-linked recessive lysosomal storage disorder caus ed by a deficiency of alpha-galactosidase A (alpha-gal; EC 3.2.1.22). In the past, it has been difficult to give an unequivocal diagnosis of carrier status in Fabry disease because of the overlap between normal and heterozygote enzyme levels. To facilitate rapid and accurate carr ier and hemizygote detection, a mutation detection strategy was devise d to determine the lesion in our Fabry disease patients. The seven alp ha-gal exons and adjacent intron boundaries from a representative memb er of each kindred were PCR amplified and analysed for the presence of sequence alterations by single-stranded conformation polymorphism (SS CP) analysis followed by PCR sequencing. Here we report the use of thi s strategy in the detection and analysis of the causative mutations in 9 patients with classic severe Fabry disease. Three deletions of 1-, 2-, and 3-bp (987delC, 717delAA, and Delta E358), five amino acid subs titutions (C52R, G128E, P205T, M284T, and N298K) and a mutation that a ffects the initiating methionine (M1I) were found in these patients. C ounting a previously reported mutation, this strategy has now successf ully detected all the Fabry disease mutations present in the 10 kindre ds that have been analysed. (C) 1996 Wiley-Liss, Inc.