摘要
目的探讨desmin基因exon6A360P错义突变与克山病心肌损伤的关系及exon6A360P错义突变是否是慢型及潜在型克山病损伤的易感基因。方法在陕西省克山病病区黄陵县店头镇和非病区西安市长安区，采用单纯随机抽样的方法，抽取年龄相匹配的慢型和潜在型克山病患者（病例组）30例及病区（内对照组）和非病区对照（外对照组）各30例。采集外周血5ml，乙二胺四乙酸（EDTA）抗凝，盐析法制备基因组DNA。采用PCR方法，用特异性的Bsp1286I内切酶对可能发生突变的exon6位点进行酶切。设内对照，在desmin基因exon4位点进行酶切，判定标准为在122bp和60bp处各出现1个条带。琼脂糖凝胶电泳分析desmin基因exon6A360P错义突变位点，判定标准为desmin基因exon6位点酶切后在184bp和66bp处各出现1个条带。结果琼脂糖凝胶电泳结果显示，内对照desmin基因exon4在Bsp1286I酶切前，在182bp处仅出现1条条带；在酶切后，被切成2个片段，在122bp和60bp处各出现1个条带。慢型及潜在型克山病患者desmin基因exon6在Bsp1286I酶切前和酶切后．均在250bp处出现了1个条带：病区和非病区对照组desmin基因酶切后．也是在250bp处出现了1个条带。3组结果相同，desmin基因exon6没有被Bsp1286I酶切开，A360P没有发生错义突变。结论慢型和潜在型克山病患者未检出desmin基因exon6A360P错义突变位点，desmin基因exon6A360P错义突变不是克山病心肌损伤的易感基因。
Objective To survey the relationship between myocardial damage of chronic and latent Keshan disease with A360P missense mutation in desmin gene exon6. Methods By clinical epidemiology method, 30 cases had been collected randomly among chronic Keshan disease patients and 30 cases equally among healthy adults （inner control groups） in Diantou town, Huangling county, Shaanxi province, a Keshan disease area, and 30 cases among health adults（outer control group） in Chang＇an district, Xian city, a normal area. Genome DNA had been extracted from 90 blood samples. Different restriction sites had been analyzed by the methods of PCR, digested by restriction endonuclease and electrophoresis. By virtue of Bsp1286 I enzyme, 122 bp and 60 bp strap could be found in control group from exon4, showing that Bsp1286 I enzyme was active. By virtue of Bsp1286 I enzyme, desmin gene exon6 was digested to 184 bp and 66 bp strip when A360P missense mutation site exited in exon6. Results One hundred and twenty two bp and 60 bp strap could be found in control group from exon4 digested by Bsp1286 I , which showed Bsp1286 I enzyme was active. Two hundred and fifty bp strips can only be found in digest products of exon6. One hundred and eighty-four and 66 bp strips which were the products of DNA digested by Bsp1286 1 could not be found. A360P missense mutation site of desmin gene exon6 had not been foundamong chronic and latent Keshan disease patients and tow control groups in Keshan disease area and in normal area. Conclusions A360P missense mutation site of desmin gene exon6 has not been found among chronic and latent Keshan disease patients. A360P missense mutation of desmin gene exon6 probably is not predisposing genes of chronic Keshan disease.
出处
《中国地方病学杂志》
CAS
CSCD
北大核心
2010年第1期46-49,共4页
Chinese Jouranl of Endemiology
