A diagnostic study on cardiomyopathy by using DNA analysis in the autopsy cases of sudden death and its application to a medicolegal judgment

方法每例心源性猝死24 h内尸检的患者取10 ml血样，冷冻于-80℃，分别取左、右心室2片固定于10%福尔马林溶液中。采用碘化钠法对血样进行解冻，并从离体外周血白细胞核中提取DNA。用一套引物对人β -肌球蛋白重链基因外显子13进行PCR扩增。pcr扩增的片段用Dde I限制性内切酶破坏，在10%聚丙烯酰胺凝胶上室温溶解。用溴化乙锭对凝胶进行染色，并在紫外光下鉴定凝胶带。采用苏木精-伊红（H-E）和azan染色对各心室切片进行组织病理学检查。结果第一例59岁男性，心脏重920 g。接下来，在一名56岁的男性中，心脏重590克。在63岁的男性中，一个人做了700克。在48岁的男性中，一个人做了530克。在27岁的男性中，一个做了450克。在65岁的男性中，一个人做了920克。25岁的男性则是680克。在48岁的男性中，一个人做了400克。在DNA分析中，分别鉴定出161个碱基对片段和129个未鉴定的碱基对片段。在心室的组织病理学检查中，我们发现心肌紊乱。

MethodsIn each case which was autopsied within 24 hours after sudden cardiac death, a 10 ml blood sample was freezed at -80ﾟC and two pieces of the left cardiac ventricle and two ones of the right ventricle were fixed in 10% formaline solution.A blod sample was defrosted and DNA was extracted from isolated nuclei of peripheral white cells by procedures of sodium iodide method. A PCR amplification of exons 13 in human beta-Myosin Heavy Chain gene was done by using a set of primers. PCR-amplified fragments were broken by Dde I restriction enzyme and resolved on a 10% polyacrylamide gel run at room temperature. The gel was stained with ethidium bromide and gel bands were identified under ultraviolet light.Each piece of the cardiac ventricle was stained with hematoxylin-eosin (H-E) and azan stains and examined histopathologically.ResultsIn the first case (59-year-old male), the heart weighed 920 g. Next, in the 56-year-old male, the heart weighed 590 g. In the 63-year-old male, the one did 700 g. In the 48-year-old male, the one did 530 g. In the 27-year-old male, the one did 450 g. In the 65-year-old male, the one did 920 g. In the 25-year-old male, the one did 680 g. In the 48-year-old male, the one did 400 g.In a DNA analysis, 161 basepair fragment was identified and 129 basepair fragment was not in each case. In histopathological study on the cardiac ventricles, we found myocardial disarray.