title: An APEX-based genotyping microarray for the screening of 168 mutations associated with familial hypercholesterolemia.
abstract: OBJECTIVE
  Familial hypercholesterolemia (FH) is an inborn disorder of lipid metabolism characterised by elevated plasma concentrations of low-density lipoprotein cholesterol and total cholesterol. This imbalance results in accelerated atherosclerosis and premature coronary heart disease. The early identification and treatment of FH patients is extremely important because it leads to significant reduction of both coronary morbidity and mortality. FH is transmitted in an autosomal dominant manner and associated predominantly with mutations in the genes encoding the low-density lipoprotein receptor (LDLR) and its ligand apolipoprotein B (APOB). To date, more than 1000 sequence variants have been described in the LDLR gene. In marked contrast to LDLR, only one APOB mutation is prevalent in Europe.
  
  
  RESULTS
  The aim of this study was, on the basis of data obtained by the molecular genetic analysis of 1945 Czech FH probands, to propose, generate, and validate a new diagnostic tool, an APEX (Arrayed Primer EXtension)-based genotyping DNA microarray called the FH chip. The FH chip contains the APOB mutation p.Arg3527Gln, all 89 LDLR point mutations and small DNA rearrangements detected in Czech FH patients, and 78 mutations frequent in other European and Asian FH populations. The validation phase revealed the sensitivity and specificity of this platform, 100% and 99.1%, respectively.
  
  
  CONCLUSIONS
  This FH chip is a rapid, reproducible, specific, and cost-effective tool for genotyping, and in combination with MLPA (multiple ligation-dependent probe amplification) represents a reliable molecular genetic protocol for the large-scale screening of FH mutations in the Czech population.
authors.affiliation: University Hospital Brno, Centre of Molecular Biology and Gene Therapy, Černopolní 9, CZ-62500 Brno, Czech Republic.
authors.forename: Lucie
authors.initials: L
authors.lastname: Dušková
authors.forename: Lenka
authors.initials: L
authors.lastname: Kopečková
authors.forename: Eva
authors.initials: E
authors.lastname: Jansová
authors.forename: Lukáš
authors.initials: L
authors.lastname: Tichý
authors.forename: Tomáš
authors.initials: T
authors.lastname: Freiberger
authors.forename: Petra
authors.initials: P
authors.lastname: Zapletalová
authors.forename: Vladimír
authors.initials: V
authors.lastname: Soška
authors.forename: Barbora
authors.initials: B
authors.lastname: Ravčuková
authors.forename: Lenka
authors.initials: L
authors.lastname: Fajkusová
mesh_terms: D001055:Apolipoproteins B / Q000235:genetics*; D018153:Czech Republic / Q000453:epidemiology; D004252:DNA Mutational Analysis; D020869:Gene Expression Profiling / Q000379:methods*; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods*; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis* / Q000453:epidemiology / Q000235:genetics; D008403:Mass Screening / Q000379:methods*; D009154:Mutation; D020411:Oligonucleotide Array Sequence Analysis; D011237:Predictive Value of Tests; D011973:Receptors, LDL / Q000235:genetics*; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity
chemical_list: D001055:Apolipoproteins B; D011973:Receptors, LDL