Isw Abstracts

Familial Hypercholesterolemia (FH) is a genetic disorder characterised by high levels of low density lipoprotein in the cardiovascular system and early onset of cardiovascular disease.FH is most commonly caused by mutations in apolipoprotein B (ApoB), low density lipoprotein receptor (LDLR), and proprotein convertase subtilisin/kexin type 9 (PCSK9) genes.FH can be treated quite effectively with lipid-lowering drugs and lifestyle changes.Genetic testing and cascade screening have been recommended by the National Institute for Health and Clinical Excellence (2008).This study reports an assay, based on a combination of multiplex PCR and biochip array hybridisation, which enables the rapid simultaneous detection of 40 mutational targets within ApoB, LDLR and PCSK9.This represents a cost-effective approach and rapid turnaround time, with easy to interpret results using the system software.The assay is based on a combination of multiplex PCR reactions and biochip array hybridisation.Innovative PCR priming permits high discrimination between multiple wild-type and mutant DNA regions which hybridise to complementary test regions on the biochip array.DNA extracted from peripheral blood and buccal cells were assayed.Using the biochip array analyser Evidence Investigator system, results are processed automatically, with analysis completed within 3 hours, from template DNA.Verification of the assay was achieved using 159 pre-characterised DNA samples in order to confirm specificity of the biochip array for detecting the FH mutations.Furthermore, validation of the array was completed using an additional set of 100 pre-characterised samples.This was completed in a blinded study showing 98% concordance.The assay detected 71% of all the point mutations in FH patients within the United Kingdom (based on 465 families from a variety of ethnic backgrounds with identified FH mutations).This rapid screening technique enables the simultaneous analysis of 40 common mutations associated with FH.This will aid in the confirmation of suspected cases of FH and in cascade screening of FH cases, hence reducing FH associated morbidity and mortality.