title: Screening and identification of familial defective apolipoprotein B-100 in clinical samples by capillary gel electrophoresis.
abstract: Familial defective apolipoprotein B-100 (FDB) is a dominantly inherited disorder. It is characterized by a decreased affinity of low density lipoprotein (LDL) for the LDL receptor, as a consequence of a substitution of adenine by guanine in exon 26 of the apolipoprotein B-100 gene, coding for the putative LDL receptor-binding domain of the mature protein. This disorder is associated with a strikingly high incidence of arteriosclerosis and tends to cause disease and premature death. In this communication we describe a rapid capillary gel electrophoretic method in combination with molecular biology techniques to facilitate the diagnosis of FDB. Mutation screening for FDB is performed by an allele-specific amplification followed by capillary gel electrophoresis (CGE). For the combined polymerase chain reaction (PCR)-CGE method, a total analysis time of only 3 h is needed, a period that is normally necessary for the run and for staining of the gel only, not including the time for PCR, gel casting, etc. In our pilot study 4 of 43 hypercholesterolemic patients were found to have the predominant apoB 3500 codon mutation. The verification is demonstrated by DNA-sequencing. This pilot study will be followed by a large cohort analysis of the south-west German population to determine the frequency of FDB in this area. The PCR-CGE method on the Dionex capillary electrophoresis system (CES I) allows rapid, fully automated detection of the mutation resulting in the unequivocal diagnosis of FDB.
authors.affiliation: Universität Tübingen, Innere Medizin Abteilung IV, Germany.
authors.forename: R
authors.initials: R
authors.lastname: Lehmann
authors.forename: M
authors.initials: M
authors.lastname: Koch
authors.forename: M
authors.initials: M
authors.lastname: Pfohl
authors.forename: W
authors.initials: W
authors.lastname: Voelter
authors.forename: H U
authors.initials: HU
authors.lastname: Häring
authors.forename: H M
authors.initials: HM
authors.lastname: Liebich
mesh_terms: D000328:Adult; D000368:Aged; D001055:Apolipoproteins B / Q000032:analysis* / Q000172:deficiency / Q000235:genetics*; D001345:Autoradiography; D001483:Base Sequence; D002624:Chemistry, Clinical / Q000379:methods; D015331:Cohort Studies; D004247:DNA / Q000235:genetics; D017931:DNA Primers / Q000737:chemistry; D019075:Electrophoresis, Capillary / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D005858:Germany; D006801:Humans; D006950:Hyperlipidemia, Familial Combined / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*; D008297:Male; D008875:Middle Aged; D010865:Pilot Projects; D016133:Polymerase Chain Reaction; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D016622:Silver Staining; D013997:Time Factors
chemical_list: D001055:Apolipoproteins B; D017931:DNA Primers; D004247:DNA