Identification of a common low density lipoprotein receptor mutation (C163Y) in the west of Scotland
Identification of a common low density lipoprotein receptor mutation (C163Y) in the west of Scotland
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DOI：
10.1136/jmg.35.7.573
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发表时间：
1998-07-01
影响因子：
4
通讯作者：
Dominiczak, AF
中科院分区：
文献类型：
--
作者：
Lee, WK;Haddad, L;Dominiczak, AF
家族性高胆固醇血症（FH）是一种常染色体共显性遗传疾病，其特征是高水平的LDL胆固醇和冠状动脉疾病的高发病率。我们的目的是跟踪低密度脂蛋白受体（LDLR）基因在个别家庭的表型FH和识别和测序的LDLR基因的突变，可能是常见的苏格兰FH人口的单链构象多态性分析（SSCP）。患者样本包括80例FH杂合子先证者，200例与先证者相关的受试者，以及另外50例正常、无关的对照受试者。通过扩增与LDLR基因座紧密连锁的19个等位基因四核苷酸微卫星来完成LDLR基因的追踪。使用LDLR基因外显子4特异性引物扩增基因组DNA，并用于SSCP分析。任何PCR产物与不同的迁移模式，通过SSCP评估，然后直接测序。除了确定一个共同的突变先证者，家庭成员进行了筛选，使用强制限制性酶切位点测定和分析，使用微板阵列对角线凝胶电泳（MADGE）。微卫星D19 S394分析在23个家系中的20个中提供了信息。在这些家族中，FH表型与LDLR基因座的分离并不矛盾。在FH先证者中，15/80例具有突变等位基因，如使用覆盖LDLR基因的整个外显子4的三对引物通过SSCP评估的。直接DNA测序显示7/15的先证者有C163 Y突变。使用PCR诱导的酶RsaI和MADGE的限制性酶切位点分析，确定了在FH先证者的家族成员中C163 Y突变与FH表型共分离。该突变等位基因在任何对照受试者中均不存在。微卫星分析已被证明可用于跟踪LDLR基因，并可与LDL胆固醇水平结合用于诊断FH，特别是在儿童和年轻人中，表型诊断可能很困难。
Familial hypercholesterolaemia (FH) is an autosomal codominant disorder characterised by high levels of LDL cholesterol and a high incidence of coronary artery disease. Our aims were to track the low density lipoprotein receptor (LDLR) gene in individual families with phenotypic FH and to identify and characterise any mutations of the LDLR gene that may be common in the west of Scotland FH population using single strand conformational polymorphism analysis (SSCP). Patient samples consisted of 80 heterozygous probands with FH, 200 subjects who were related to the probands, and a further 50 normal, unrelated control subjects. Tracking of the LDLR gene was accomplished by amplification of a 19 allele tetranucleotide microsatellite that is tightly Linked to the LDLR gene locus. Primers specific for exon 4 of the LDLR gene were used to amplify genomic DNA and used for SSCP analysis. Any PCR products with different migration patterns as assessed by SSCP were then sequenced directly In addition to identifying probands with a common mutation, family members were screened using a forced restriction site assay and analysed using microplate array diagonal gel electrophoresis (MADGE). Microsatellite D19S394 analysis was informative in 20 of 23 families studied. In these families there was no inconsistency with segregation of the FH phenotype with the LDLR locus. Of the FH probands, 15/80 had a mutant allele as assessed by SSCP using three pairs of primers covering the whole of exon 4 of the LDLR gene. Direct DNA sequencing showed that 7/15 of the probands had a C163Y mutation. Using a PCR induced restriction site assay for the enzyme RsaI and MADGE, it was determined that the C163Y mutation cosegregated with the FH phenotype in family members of the FH probands. This mutant allele was not present in any of the control subjects. Microsatellite analysis has proven useful in tracking the LDLR gene and could be used in conjunction with LDL cholesterol levels to diagnose FH, especially in children and young adults where phenotypic diagnosis can be difficult.