The Ivestigation Of Cardiac Myosin Bindiong Protein-C Gene Mutation In Pations With Hypertrophic Cardiomyopathy

1. To investigate the situation of Cardiac myosin binding protein-C gene mutation in Chinese population of Shandong province with hypertrophic cardiomyopathy(HCM); 2. To research if there are differences between the Chinese people and foreigners in the incidence of Cardiac myosin binding protein-C gene mutation in patients with hypertrophic cardiomyopathy; 3. To analysis the correlation between genotypes and phenotypes; 4.To establish a method to diagnose hypertrophic cardiomyopathy preclinically at gene level.Methods: 1. Ninety-two unrelated clinical patients with HCM were chosen for the study. After informed consent a venous blood sample for extraction of DNA was obtained. In addition, one hundred healthy individuals were examined as normal controls; 2. High molecular weight DNA was extracted from peripheral blood lymphocytes of patients and controls with the SDS proteinase K method and phenol/chloroform extraction; 3. The primers used for amplication of exonl4-15,17, 25, 27, 33of Cardiac myosin binding protein-C gene were from adjacent intronic sequence. Protein coding exons 14-15,17, 25, 27, 33of Cardiac myosin binding protein-C was amplified using polymerase chain reaction(PCR). The PCR products were examined by means of agar-gel electrophoresis to assay the quantity and specialty; 4. Single strand conformation polymorphism(SSCP) method was used to detect possible gene mutations in the PCR products. Different conditions were used to enhance the sensitivityof the method including different polypropylene concention, different propotion of PCR products and cushion fluid etc; 5.Sequence the suspicious PCR products for exact mutant point. Perform clinical studies and examine the incidence of sudden cardiac death within the family.Results: 1. We succeeded in amplifying the five exons of the ninety two patients. 2. All of the PCR products were examined by SSCP method through different conditions and vivid single strand belts have been got. 3. By contrast with normal controls, we have found a missence mutation T445S for the first time in a HCM patient. 4.By contrast to the other HCM patients, we couldn' t find the paticurity of the patient harboring the mutation.Conclusions: 1. We have found a misence mutation for the first time in a HCM patient. 2. The incidence of mutations in gene encoding Cardiac myosin binding protein-C in HCM patients of Chinese population is much lower than what have been previously reported. 3. The genotypes and phenotypes are not simply in according with each other. 4.There is a long way to go to conquer this disease.