title: Simultaneous detection of multiple familial hypercholesterolemia mutations facilitates an improved diagnostic service in South african patients at high risk of cardiovascular disease.
abstract: OBJECTIVE
  DNA testing can provide a definitive diagnosis of familial hypercholesterolemia (FH), even in the absence of the clinical characteristics of this inherited cardiovascular disease (CVD) subtype. Our aim was to design a rapid diagnostic assay capable of simultaneously analyzing seven point mutations in the low-density lipoprotein receptor (LDLR) gene, which occur at high frequency in South African FH patients.
  
  
  METHODS
  The test is based on multiplex DNA amplification and hybridization to membrane strips presenting a parallel array of immobilized allele-specific oligonucleotide probes.
  
  
  RESULTS
  A reverse-hybridization assay for genotyping LDLR point mutations D154N, D200G, D206E, C356Y, G361V, V408M, and P664L was set-up and validated using pretyped human DNA samples, as well as recombinant plasmid clones containing mutant alleles. The procedure is rapid (6 hours) and may be automated to a large extent.
  
  
  CONCLUSIONS
  The new FH strip-assay forms an important part of the comprehensive cardiovascular genetic screen offered routinely to high-risk population groups in South Africa. A genetic approach based on FH testing in conjunction with other 'genetic' CVD risk factors is feasible and justified, since the spectrum of disease-related mutations have been defined to a large extent in the genetically distinct population groups of South Africa. Knowledge of a significantly increased CVD risk due to the presence of gene variations, which can be targeted for risk reduction by the avoidance of relevant environmental risk factors and the appropriate treatment, provides a powerful message to motivate people into implementing preventative measures based on their genetic profile.
authors.affiliation: Division of Human Genetics, Faculty of Heath Sciences, University of Stellenbosch, Tygerberg, and Genecare Molecular Genetics (Pty) Ltd, Christiaan Barnard Memorial Hospital, Cape Town, South Africa. mjk@genecare.co.za
authors.forename: Maritha J
authors.initials: MJ
authors.lastname: Kotze
authors.forename: Gernot
authors.initials: G
authors.lastname: Kriegshäuser
authors.forename: Rochelle
authors.initials: R
authors.lastname: Thiart
authors.forename: Nico J P
authors.initials: NJ
authors.lastname: de Villiers
authors.forename: Charlotte L
authors.initials: CL
authors.lastname: Scholtz
authors.forename: Fritz
authors.initials: F
authors.lastname: Kury
authors.forename: Anne
authors.initials: A
authors.lastname: Moritz
authors.forename: Christian
authors.initials: C
authors.lastname: Oberkanins
mesh_terms: D019943:Amino Acid Substitution; D002318:Cardiovascular Diseases / Q000175:diagnosis / Q000453:epidemiology / Q000235:genetics*; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000235:genetics*; D017354:Point Mutation / Q000235:genetics*; D016133:Polymerase Chain Reaction / Q000379:methods; D011973:Receptors, LDL / Q000235:genetics*; D012307:Risk Factors; D013019:South Africa / Q000453:epidemiology
chemical_list: D011973:Receptors, LDL