title: Rapid diagnosis of familial defective apolipoprotein B-100.
abstract: A method is described for the rapid, economic and non-radioactive examination of DNA samples from hypercholesterolaemic patients for familial defective apolipoprotein B-100, using a modified polymerase chain reaction (PCR) protocol and restriction enzyme isoform genotyping. Because of the high prevalence of familial defective apolipoprotein B-100, which is estimated to be one in 500 in most screened general populations, interest is focussed on a simple method for detection of this point mutation. In our protocol a diagnostic restriction site is created by PCR, using a specifically designed partly mismatched primer. In the case of familial defective apolipoprotein B-100 the amplified DNA segment contains an additional ScaI site, whereas DNA amplified from the normal allele is resistant to ScaI digestion. A rapid differentiation between the two alleles is achieved by agarose gel electrophoresis of the ScaI-digested PCR product.
authors.affiliation: Institut für Klinische Chemie, Universität zu Köln.
authors.forename: J
authors.initials: J
authors.lastname: Geisel
authors.forename: T
authors.initials: T
authors.lastname: Schleifenbaum
authors.forename: B
authors.initials: B
authors.lastname: Weisshaar
authors.forename: K
authors.initials: K
authors.lastname: Oette
mesh_terms: D053299:Apolipoprotein B-100; D001055:Apolipoproteins B / Q000235:genetics*; D001483:Base Sequence; D008078:Cholesterol, LDL / Q000097:blood*; D003062:Codon / Q000235:genetics; D005260:Female; D006801:Humans; D006937:Hypercholesterolemia / Q000097:blood / Q000235:genetics*; D007962:Leukocytes / Q000502:physiology; D008297:Male; D008969:Molecular Sequence Data; D010375:Pedigree; D016133:Polymerase Chain Reaction / Q000379:methods; D015183:Restriction Mapping
chemical_list: D053299:Apolipoprotein B-100; D001055:Apolipoproteins B; D008078:Cholesterol, LDL; D003062:Codon