Equal contribution as senior and corresponding authors.
Genetic diagnosis of familial hypercholesterolaemia by targeted next-generation sequencing
Article first published online: 20 MAY 2014
© 2014 The Authors. Journal of Internal Medicine published by John Wiley & Sons Ltd on behalf of The Association for the Publication of the Journal of Internal Medicine.
This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.
Journal of Internal Medicine
Volume 276, Issue 4, pages 396–403, October 2014
How to Cite
Genetic diagnosis of familial hypercholesterolaemia by targeted next-generation sequencing. J Intern Med 2014; 276: 396–403., , , , , , , , , (University of Gothenburg, Gothenburg, Sweden; University Magna Graecia of Catanzaro, Italy; University of Milan, Italy; Progenika Biopharma SA, Derio, Spain).
- Issue published online: 20 SEP 2014
- Article first published online: 20 MAY 2014
- Accepted manuscript online: 1 MAY 2014 09:56PM EST
- Swedish Research Council
- Swedish Diabetes Foundation
- Swedish Heart-Lung Foundation
- Region Västra Götaland
- Sahlgrenska University Hospital
- Wilhelm and Martina Lundgren Science Fund
- Fysiografiska Sällskapet
- European Commission
- European Social Fund
- Calabria Region
- familial hypercholesterolaemia;
- genetic diagnosis;
- LDL receptor;
- next-generation sequencing;
The aim of this study was to combine clinical criteria and next-generation sequencing (pyrosequencing) to establish a diagnosis of familial hypercholesterolaemia (FH).
Design, setting and subjects
A total of 77 subjects with a Dutch Lipid Clinic Network score of ≥3 (possible, probable or definite FH clinical diagnosis) were recruited from the Lipid Clinic at Sahlgrenska Hospital, Gothenburg, Sweden. Next-generation sequencing was performed in all subjects using SEQPRO LIPO RS, a kit that detects mutations in the low-density lipoprotein receptor (LDLR), apolipoprotein B (APOB), proprotein convertase subtilisin/kexin type 9 (PCSK9) and LDLR adapter protein 1 (LDLRAP1) genes; copy-number variations in the LDLR gene were also examined.
A total of 26 mutations were detected in 50 subjects (65% success rate). Amongst these, 23 mutations were in the LDLR gene, two in the APOB gene and one in the PCSK9 gene. Four mutations with unknown pathogenicity were detected in LDLR. Of these, three mutations (Gly505Asp, Ile585Thr and Gln660Arg) have been previously reported in subjects with FH, but their pathogenicity has not been proved. The fourth, a mutation in LDLR affecting a splicing site (exon 6–intron 6) has not previously been reported; it was found to segregate with high cholesterol levels in the family of the proband.
Using a combination of clinical criteria and targeted next-generation sequencing, we have achieved FH diagnosis with a high success rate. Furthermore, we identified a new splicing-site mutation in the LDLR gene.