中国地方病学杂志
中國地方病學雜誌
중국지방병학잡지
CHINESE JOURNAL OF ENDEMIOLOGY
2010年
1期
46-49
,共4页
克山病%基因%desmin%点突变
剋山病%基因%desmin%點突變
극산병%기인%desmin%점돌변
Keshan disease%Gene,desmin%Point mutation
目的 探讨desmin基因exon6 A360P错义突变与克山病心肌损伤的关系及exon6 A360P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5 ml,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Bsp1286 Ⅰ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122 bp和60 bp处各出现1个条带.琼脂糖凝胶电泳分析desmin基因exon6 A360P错义突变位点,判定标准为desmin基因exon6位点酶切后在184 bp和66 bp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Bsp1286 Ⅰ酶切前,在182 bp处仅出现1条条带;在酶切后,被切成2个片段,在122 bp和60 bp处各出现1个条带.慢型及潜在型克山病患者desmin基因exon6在Bsp1286 Ⅰ酶切前和酶切后,均在250 bp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250 bp处出现了1个条带.3组结果相同,desmin基因exon6没有被Bsp1286 Ⅰ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exon6 A360P错义突变位点,desmin基因exon6 A360P错义突变不是克山病心肌损伤的易感基因.
目的 探討desmin基因exon6 A360P錯義突變與剋山病心肌損傷的關繫及exon6 A360P錯義突變是否是慢型及潛在型剋山病損傷的易感基因.方法 在陝西省剋山病病區黃陵縣店頭鎮和非病區西安市長安區,採用單純隨機抽樣的方法,抽取年齡相匹配的慢型和潛在型剋山病患者(病例組)30例及病區(內對照組)和非病區對照(外對照組)各30例.採集外週血5 ml,乙二胺四乙痠(EDTA)抗凝,鹽析法製備基因組DNA.採用PCR方法,用特異性的Bsp1286 Ⅰ內切酶對可能髮生突變的exon6位點進行酶切.設內對照,在desmin基因exon4位點進行酶切,判定標準為在122 bp和60 bp處各齣現1箇條帶.瓊脂糖凝膠電泳分析desmin基因exon6 A360P錯義突變位點,判定標準為desmin基因exon6位點酶切後在184 bp和66 bp處各齣現1箇條帶.結果 瓊脂糖凝膠電泳結果顯示,內對照desmin基因exon4在Bsp1286 Ⅰ酶切前,在182 bp處僅齣現1條條帶;在酶切後,被切成2箇片段,在122 bp和60 bp處各齣現1箇條帶.慢型及潛在型剋山病患者desmin基因exon6在Bsp1286 Ⅰ酶切前和酶切後,均在250 bp處齣現瞭1箇條帶;病區和非病區對照組desmin基因酶切後,也是在250 bp處齣現瞭1箇條帶.3組結果相同,desmin基因exon6沒有被Bsp1286 Ⅰ酶切開,A360P沒有髮生錯義突變.結論 慢型和潛在型剋山病患者未檢齣desmin基因exon6 A360P錯義突變位點,desmin基因exon6 A360P錯義突變不是剋山病心肌損傷的易感基因.
목적 탐토desmin기인exon6 A360P착의돌변여극산병심기손상적관계급exon6 A360P착의돌변시부시만형급잠재형극산병손상적역감기인.방법 재합서성극산병병구황릉현점두진화비병구서안시장안구,채용단순수궤추양적방법,추취년령상필배적만형화잠재형극산병환자(병례조)30례급병구(내대조조)화비병구대조(외대조조)각30례.채집외주혈5 ml,을이알사을산(EDTA)항응,염석법제비기인조DNA.채용PCR방법,용특이성적Bsp1286 Ⅰ내절매대가능발생돌변적exon6위점진행매절.설내대조,재desmin기인exon4위점진행매절,판정표준위재122 bp화60 bp처각출현1개조대.경지당응효전영분석desmin기인exon6 A360P착의돌변위점,판정표준위desmin기인exon6위점매절후재184 bp화66 bp처각출현1개조대.결과 경지당응효전영결과현시,내대조desmin기인exon4재Bsp1286 Ⅰ매절전,재182 bp처부출현1조조대;재매절후,피절성2개편단,재122 bp화60 bp처각출현1개조대.만형급잠재형극산병환자desmin기인exon6재Bsp1286 Ⅰ매절전화매절후,균재250 bp처출현료1개조대;병구화비병구대조조desmin기인매절후,야시재250 bp처출현료1개조대.3조결과상동,desmin기인exon6몰유피Bsp1286 Ⅰ매절개,A360P몰유발생착의돌변.결론 만형화잠재형극산병환자미검출desmin기인exon6 A360P착의돌변위점,desmin기인exon6 A360P착의돌변불시극산병심기손상적역감기인.
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 exnn4,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 Ⅰ could not be found. A360P missense mutation site of desmin gene exon6 had not been found among 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.
