Molecular testing for familial hypercholesterolaemia-associated mutations in a UK-based cohort: Development of an NGS-based method and comparison with multiplex polymerase chain reaction and oligonucleotide arrays

BackgroundDetection of disease-associated mutations in patients with familial hypercholesterolaemia is crucial for early interventions to reduce risk of cardiovascular disease. Screening for these mutations represents a methodological challenge since more than 1200 different causal mutations in the low-density lipoproteinreceptor has been identified. A number of methodological approaches have been developed for screening by clinical diagnostic laboratories.MethodsUsing primers targeting, the low-density lipoproteinreceptor, apolipoprotein B, and proprotein convertase subtilisin/ kexintype 9, we developed a novel Ion Torrent-based targeted re-sequencing method. We validated this in a West Midlands-UK small cohort of 58 patients screened in parallel with other mutation-targeting methods, such as multiplex polymerase chain reaction(Elucigene FH20), oligonucleotide arrays (Randox familial hypercholesterolaemia array) or the Illumina next-generation sequencing platform.ResultsIn this small cohort, the next-...