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Screening and identification of familial defective apolipoprotein B-100 in clinical samples by capillary gel electrophoresis.

Authors :
Lehmann R
Koch M
Pfohl M
Voelter W
Häring HU
Liebich HM
Source :
Journal of chromatography. A [J Chromatogr A] 1996 Sep 13; Vol. 744 (1-2), pp. 187-94.
Publication Year :
1996

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.

Details

Language :
English
ISSN :
0021-9673
Volume :
744
Issue :
1-2
Database :
MEDLINE
Journal :
Journal of chromatography. A
Publication Type :
Academic Journal
Accession number :
8843667
Full Text :
https://doi.org/10.1016/0021-9673(96)00244-0