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Genetic diagnosis of familial hypercholesterolaemia by targeted next‐generation sequencing
Author(s) -
Maglio C.,
Mancina R. M.,
Motta B. M.,
Stef M.,
Pirazzi C.,
Palacios L.,
Askaryar N.,
Borén J.,
Wiklund O.,
Romeo S.
Publication year - 2014
Publication title -
journal of internal medicine
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.625
H-Index - 160
eISSN - 1365-2796
pISSN - 0954-6820
DOI - 10.1111/joim.12263
Subject(s) - pcsk9 , ldl receptor , familial hypercholesterolemia , kexin , apolipoprotein b , medicine , exon , genetics , proband , mutation , gene mutation , gene , bioinformatics , biology , lipoprotein , cholesterol
Objectives 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 ( PCSK 9 ) and LDLR adapter protein 1 ( LDLRAP 1) genes; copy‐number variations in the LDLR gene were also examined. Results 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 PCSK 9 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. Conclusions 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.