家族性致心律失常性右室心肌病与微卫星遗传标志的连锁分析

Linkage analysis of several microsatellite genetic markers with arrhythmogenic right ventricular cardiomyopathy in 5 Chinese families

目的 研究探索致心律失常性右室心肌病 (ARVC)家系与 3个微卫星遗传标志的连锁关系 ,进行ARVC的基因定位。方法 选择微卫星遗传标志D2S15 2、D14S2 5 2和D10S16 6 4 ,对 12 1例背景人群和 5个中国人ARVC家系 (家系编号 1～ 5 ) ,用每个微卫星引物扩增家系和背景人群DNA的微卫星片段 ,在DNA手工测序电泳仪槽上进行恒功率变性聚丙烯酰胺凝胶电泳、银染、读取等位基因片段 ,在常染色体显性和隐性两种遗传模式下进行连锁分析。结果 (1)根据D2S15 2的连锁资料 ,在常染色体显性遗传模式下 ,1～ 5号家系的连锁值 (Logarithmofodd ,LOD值 )分别为 2 17、- 0 5 9、-∞ (负无穷大 )、- (表示此遗传模式下不支持连锁分析 )、-∞ ,均为θ =0。在常染色体隐性遗传模式下 ,1～ 5号家系的LOD值分别为 -∞、-∞、-∞、-∞、0 18。 (2 )根据D14S2 5 2的连锁资料 ,在常染色体显性遗传模式下 ,1～ 5号家系的LOD值分别为 -、-、-∞、-、0。在常染色体隐性遗传模式下 ,1～ 5号家系的LOD值分别为 -∞、- 0 81、-∞、-∞、0 0 9。 (3)根据D10S16 6 4的连锁资料 ,在常染色体显性遗传模式下 ,1～ 5号家系的LOD值分别为 -、-、0 5 4、-、0 6 0。在常染色体隐性遗传模式下 ,1～ 5号家系的LOD值分别为 -、-∞、-∞、-∞、

Objective The mechanism of arrhythmogenic right ventricular cardiomyopathy (ARVC) is not clear and the genetic occurrence rate is up to 44%. Finding the causative ARVC gene is critical to exploring this mechanism. This study explored the linkage relationship between the selected microsatellite genetic markers and ARVC in 5 Chinese families. Methods The microsatellite genetic markers D2S152, D14S252 and D10S1664, 5 Chinese families(Family No.1 - 5)and a population of 121 persons were selected. The polymorphic repeat regions of the markers were amplified directly from genomic DNA and assessed on polyacrylamide sequencing gels (4%). The samples of the PCR products were electrophoresed (electric constant power supply: ECPS 3000/150, Pharmacia) on 4% denaturing polyacrylamide gels (acrylamide: Bis=39∶1) for 1 hour or so at the stable power of 35W. Gels were silver-stained using the protocol provided by Promega. The classical linkage analysis was performed under the genetic modes of autosomal dominant and recessive. Results (1) The Logarithm of odd (LOD) scores of D2S152 with ARVC in No.1 - 5 family were 2.17, -0.59, -∞, -( indicating that linkage can not be performed in this mode),-∞ respectively in autosomal dominant mode (θ=0, respectively )and were -∞,-∞, -∞, -∞, 0.18 respectively in the autosomal recessive mode. (2) The LOD scores of D14S252 with ARVC in No.1 - 5 family were -, -, -∞, -, 0 respectively in autosomal dominant mode and were -∞ , -0.81, -∞, -∞, 0.09 respectively in autosomal recessive mode. (3) The LOD scores of D10S1664 with ARVC in No.1-5 family were -, -, 0.54, -, 0.60 respectively in autosomal dominant mode and were -, -∞, -∞, -∞, -∞ respectively in autosomal recessive mode. Conclusions (1) The LOD score in No.1 family with D2S152 was 2.17, which means that the rate of linkage: no linkage is 150∶1. This suggests that the possible causative ARVC gene near this marker exist. The linkage relationship with D2S152 was not found in No.3 family and more information is needed about No. 2, 4 and 5 families so that the linkage relation can be further investigated. (2) The linkage relationship with D14S252 was not found in the No.3 family and more information is needed about No.1,2, 4 and 5 families. (3) The linkage relationship with D10S1664 was not found in all 5 families and also more information is needed.