title: Rapid screening for specific mutations in patients with a clinical diagnosis of familial hypercholesterolaemia.
abstract: We describe a rapid screening procedure to identify known DNA sequence changes in individuals diagnosed as having heterozygous familial hypercholesterolaemia (FH). The screening is made possible by combining a rapid DNA extraction protocol and small scale polymerase chain reaction DNA amplification, followed by oligonucleotide melting or restriction enzyme digestion. We have screened for two different mutations; firstly a mutation in the apolipoprotein B (apo B) gene that results in the substitution of glutamine (Gln) for arginine (Arg) at amino acid residue 3500 (apo B3500 mutation). Apo B is the principal component of the protein moiety of low density lipoprotein (LDL) and the mutation reduces the affinity for the LDL receptor (LDL-R). Secondly we have screened for a point mutation in the LDL-R gene itself that creates a new Pst I restriction enzyme site. This mutation in the LDL-R gene (LDL-R664 mutation) results in the substitution of leucine (Leu) for proline (Pro) at amino acid 664 and is known to slow processing of the LDL-R precursor to the mature form and to reduce the affinity of the receptor on the cell surface for LDL. In 77 unrelated patients with a clinical diagnosis of FH two out of 77 (2.6%) were positive for the apo B3500 mutation. Three (3.9%) were positive for the LDL-R664 mutation. Thus these two mutations might account for 5-6% of patients in the U.K. with a clinical diagnosis of FH (5000-6000 people).(ABSTRACT TRUNCATED AT 250 WORDS)
authors.affiliation: Charing Cross Sunley Research Centre, London, U.K.
authors.forename: P
authors.initials: P
authors.lastname: Talmud
authors.forename: A
authors.initials: A
authors.lastname: Tybjaerg-Hansen
authors.forename: D
authors.initials: D
authors.lastname: Bhatnagar
authors.forename: A
authors.initials: A
authors.lastname: Mbewu
authors.forename: J P
authors.initials: JP
authors.lastname: Miller
authors.forename: P
authors.initials: P
authors.lastname: Durrington
authors.forename: S
authors.initials: S
authors.lastname: Humphries
mesh_terms: D000328:Adult; D001055:Apolipoproteins B / Q000235:genetics; D005260:Female; D005820:Genetic Testing / Q000379:methods*; D006579:Heterozygote; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis* / Q000235:genetics; D008297:Male; D008875:Middle Aged; D009154:Mutation; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D011973:Receptors, LDL / Q000235:genetics
chemical_list: D001055:Apolipoproteins B; D015345:Oligonucleotide Probes; D011973:Receptors, LDL