Blanch L C, Meaney C, Morris C P
Department of Chemical Pathology, Women's and Children's Hospital, North Adelaide, S.A., Australia.
Hum Mutat. 1996;8(1):38-43. doi: 10.1002/(SICI)1098-1004(1996)8:1<38::AID-HUMU5>3.0.CO;2-L.
Fabry disease is an X-linked recessive lysosomal storage disorder caused 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 carrier and hemizygote detection, a mutation detection strategy was devised to determine the lesion in our Fabry disease patients. The seven alpha-gal exons and adjacent intron boundaries from a representative member of each kindred were PCR amplified and analysed for the presence of sequence alterations by single-stranded conformation polymorphism (SSCP) analysis followed by PCR sequencing. Here we report the use of this 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 substitutions (C52R, G128E, P205T, M284T, and N298K) and a mutation that affects the initiating methionine (M1I) were found in these patients. Counting a previously reported mutation, this strategy has now successfully detected all the Fabry disease mutations present in the 10 kindreds that have been analysed.
法布里病是一种X连锁隐性溶酶体贮积症，由α-半乳糖苷酶A（α-gal；EC 3.2.1.22）缺乏引起。过去，由于正常水平与杂合子酶水平存在重叠，很难对法布里病携带者状态做出明确诊断。为便于快速、准确地检测携带者和半合子，设计了一种突变检测策略来确定我们法布里病患者的病变情况。对每个家系的一名代表性成员的7个α-gal外显子及相邻内含子边界进行PCR扩增，通过单链构象多态性（SSCP）分析，随后进行PCR测序，分析序列改变的存在情况。在此，我们报告该策略在9例经典重型法布里病患者致病突变检测和分析中的应用。在这些患者中发现了3个分别缺失1个、2个和3个碱基对的缺失突变（987delC、717delAA和delta E358）、5个氨基酸替代突变（C52R、G128E、P205T、M284T和N298K）以及1个影响起始甲硫氨酸的突变（M1I）。算上之前报道的一个突变，该策略现已成功检测出在已分析的10个家系中存在的所有法布里病突变。
