The Ivestigation Of Mutations In Patients With Anderson Fabry Disease

Objective: 1.By screening the Anderson fabry patients with molecular biological methods in patients who have been diagnosed as hypertrophic cardiomyopahty , we investigate the probability of misdiagnosing Anderson fabry disease as hypertrophic cardiomyopathy ; 2. To find mutations which can lead to Anderson fabry disease; 3. To research if there are differences between the Chinese people and foreigners in the incidence and clinical manifestation of Anderson fabry disease; 4.To analysis the correlation between the genotype and phenotype; 5.To establish a method to diagnose Anderson fabry preclinically at gene level.Methods: 1. 102 unrelated clinical patients diagnosed as hypertrophiccardiomyopathy were chosen for the study. After informed consent, a venousblood sample for extraction of DNA was obtained. In addition, one hundredhealthy individuals were examined as normal controls. Plasma DNA wasextracted from peripheral blood lymphocytes of patients and controls withthe SDS proteinase K method and phenol/chloroform extraction; 2. Theprimers used for amplication of seven exons of Cardiac GLA gene were fromadjacent intronic sequence. Seven exons of Cardiac GLA gene wereamplified using polymerase chain reaction(PCR). The PCR products wereexamined by means of 1. 5% agar-gel electrophoresis to assay the quantityand specialty; 3. Single strand conformation polymorphism(SSCP) was usedto detect possible gene mutations in the PCR products. Differentconditions were used to enhance the sensitivity of the method; 4. Sequencethe seven exons and adjacent intronic sequence of patients with changedstrap for exact mutant point. 5. The a-galactosidase enzyme activity of...