Genetic diagnosis of familial hypercholesterolaemia using a rapid biochip array assay for 40 common LDLR, APOB and PCSK9 mutations.
Familial hypercholesterolaemia (FH) leads to a lifelong increase in plasma LDL levels with subsequent increase in premature vascular disease. Early diagnosis and treatment is the key to effective management of this condition. This research aims to produce a simple and cost effective genetic test which could identify the majority (71%) of mutations causing FH in the UK and Ireland.



The Randox Biochip Array Technology was used to detect 40 point mutations in LDLR, APOB and PCSK9 genes, over two 5 × 5 arrays. This technology uses multiplex allele specific PCR and biochip array hybridisation, followed by a chemiluminescence detection system and software for automated mutation calling.



The FH biochip array assay was validated in the Belfast Genetics Laboratory using 199 cascade screening samples previously sequenced for known FH causing family mutations, the overall sensitivity was 98%. The assay was then used for routine testing of 663 patients with possible FH, from clinics across the UK and Ireland. A total of 49 (7.4%) mutation positive individuals were identified, however, for the clinics in England the detection rate was 12.9%. Further analysis of 120 biochip negative patients, using DNA sequencing, did not identify any false negatives.



The FH biochip array provides a rapid and reliable genetic test for the majority of FH causing point mutations in the UK and Ireland. A total of 32 samples can be run in 3 h. This allows clinics to evaluate additional patients for a possible diagnosis of FH such as patients with high LDL, patients with early onset coronary disease, and patients with relatives known to have FH.
Northern Ireland Regional Genetics Centre, Belfast City Hospital, Belfast Health and Social Care Trust, Belfast, Northern Ireland, UK.
Rosalind
R
Martin
Randox Laboratories Ltd., Crumlin, Northern Ireland, UK.
Mark
M
Latten
Northern Ireland Regional Genetics Centre, Belfast City Hospital, Belfast Health and Social Care Trust, Belfast, Northern Ireland, UK.
Padraig
P
Hart
Randox Laboratories Ltd., Crumlin, Northern Ireland, UK.
Helena
H
Murray
Northern Ireland Regional Genetics Centre, Belfast City Hospital, Belfast Health and Social Care Trust, Belfast, Northern Ireland, UK.
Deborah A
DA
Bailie
Randox Laboratories Ltd., Crumlin, Northern Ireland, UK.
Martin
M
Crockard
Randox Laboratories Ltd., Crumlin, Northern Ireland, UK.
John
J
Lamont
Randox Laboratories Ltd., Crumlin, Northern Ireland, UK.
Peter
P
Fitzgerald
Northern Ireland Regional Genetics Centre, Belfast City Hospital, Belfast Health and Social Care Trust, Belfast, Northern Ireland, UK; Randox Laboratories Ltd., Crumlin, Northern Ireland, UK; Centre for Public Health, Queens University, Belfast, Northern Ireland, UK. Electronic address: colin.graham@randox.com.
Colin A
CA
Graham
D000483:Alleles; D053299:Apolipoprotein B-100 / Q000235:genetics*; D004252:DNA Mutational Analysis; D004739:England; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000175:diagnosis* / Q000235:genetics; D007494:Ireland; D009660:Northern Ireland; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods*; D017354:Point Mutation; D000071449:Proprotein Convertase 9 / Q000235:genetics*; D011973:Receptors, LDL / Q000235:genetics*; D015203:Reproducibility of Results; D017422:Sequence Analysis, DNA; D016896:Treatment Outcome; D014852:Wales
C569471:APOB protein, human; D053299:Apolipoprotein B-100; C573412:LDLR protein, human; D011973:Receptors, LDL; C472125:PCSK9 protein, human; D000071449:Proprotein Convertase 9
Biochip array; Familial hypercholesterolaemia; Genetic diagnosis; Mutation detection