Molecular Genetic Diagnosis of a Family with Hypercholesterolemia by a Mismatched PCR-RFLP Method for Genotyping Single Base Substitution of the LDL Receptor Gene.

Abstract

Plasma lipid and lipoprotein levels reflect in part the influence of relevant genetic loci. Defects at some of these loci account for specific types of dyslipoproteinemia occurring with regularity among family members. In the course of familial investigations of coronary artery disease, we identified an family in which several members were affected with elevated low density lipoprotein (LDL) cholesterol levels. To study the genetic defects responsible for plasma lipoprotein abnormality in this pedigree, we developed a simple method for genotyping a single base substitution that does not affect a restriction recognition enzyme site in exon 10 of the LDL receptor gene. Using our mismatched PCR method, this G->A substitution at nucleotide 1413 could be genotyped in the form of a biallelic restriction fragment length polymorphism (RFLP) after digestion with restriction enzyme Hpa II. Linkage analysis using this molecular method demonstrated that the defect at the LDL receptor locus is responsible for elevated LDL cholesterol phenotype observed in this family by segregation of defective alleles at the LDL receptor locus with the disease (peak decimal logarithm of odds score>3.0).

High‐throughput single nucleotide polymorphism typing by fluorescent single‐strand conformation polymorphism analysis with capillary electrophoresis

“…This polymorphism was first described as a SSCP variant which was very difficult to type [19]. Then, Yamaki et al [20] developed a mismatched RFLP method for the typing of LDLR G1413A. However, in the present study, we successfully typed this SNP by CE-SSCP analysis.…”

Association of low-density lipoprotein receptor polymorphisms and outcome of hepatitis C infection

“…However, it has been shown that the G allele of this SNP cosegregated with elevated LDL cholesterol without exception in a Japanese pedigree. 35 The silent change in exon 12 has been shown to be associated with altered concentrations of total and LDL cholesterol in FH studies. 31,33 Another study showed that this SNP was associated with obesity overall, but not with plasma lipid levels.…”