Array-based resequencing for mutations causing familial hypercholesterolemia

Kuan Rau Chiou, Min Ji Charng, Hua Mei Chang

研究成果: 雜誌貢獻文章

20 引文 (Scopus)

摘要

Background: Familial hypercholesterolemia (FH) is a heterogeneous autosomal dominant disease with a prevalence of 1 in 500. To date, over 1200 unique pathogenic mutations have been identified in at least 3 genes. The large allelic and genetic heterogeneity of FH requires high-throughput, rapid, and affordable mutation detection technology to efficiently integrate molecular screening into clinical practice. We developed an array-based resequencing assay to facilitate genetic testing in FH patients. Methods and results: We designed a custom DNA resequencing array to detect mutations on all 3 FH-causing genes - LDL receptor (LDLR), apolipoprotein B (APOB), and proprotein convertase subtilisin/kexin type 9 gene (PCSK9) - and 290 known insertion/deletion mutations on LDLR. We verified FH array performance by analyzing 35 previously sequenced subjects (21 with point mutations, 2 insertions, 7 deletions, and 5 healthy controls) and blindly screening 125 FH patients. The average microarray call rate was 98.45% and the agreement between microarray and capillary sequencing was 99.99%. The FH array detected mutations by using automated software analysis, followed by manual review in 28 of the 30 subjects (pickup rate, 93.3%). In the blinded study, the FH array detected at least 1 mutation in 77.5% of patients clinically diagnosed with definite FH according to Simon Broome FH criteria and in 52.9% with probable FH diagnosis. Conclusions: The high-throughput FH resequencing array detects LDLR, APOB, and PCSK9 with high efficiency and accuracy and identifies disease-causing mutations. Thus, it facilitates large-scale screening of the heterogeneous FH populations.
原文英語
頁(從 - 到)383-389
頁數7
期刊Atherosclerosis
216
發行號2
DOIs
出版狀態已發佈 - 六月 1 2011
對外發佈Yes

指紋

Hyperlipoproteinemia Type II
Mutation
LDL Receptors
Apolipoproteins B
Genes
INDEL Mutation
Genetic Heterogeneity
Genetic Testing
Oligonucleotide Array Sequence Analysis
Point Mutation

ASJC Scopus subject areas

  • Cardiology and Cardiovascular Medicine

引用此文

Array-based resequencing for mutations causing familial hypercholesterolemia. / Chiou, Kuan Rau; Charng, Min Ji; Chang, Hua Mei.

於: Atherosclerosis, 卷 216, 編號 2, 01.06.2011, p. 383-389.

研究成果: 雜誌貢獻文章

Chiou, Kuan Rau ; Charng, Min Ji ; Chang, Hua Mei. / Array-based resequencing for mutations causing familial hypercholesterolemia. 於: Atherosclerosis. 2011 ; 卷 216, 編號 2. 頁 383-389.
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abstract = "Background: Familial hypercholesterolemia (FH) is a heterogeneous autosomal dominant disease with a prevalence of 1 in 500. To date, over 1200 unique pathogenic mutations have been identified in at least 3 genes. The large allelic and genetic heterogeneity of FH requires high-throughput, rapid, and affordable mutation detection technology to efficiently integrate molecular screening into clinical practice. We developed an array-based resequencing assay to facilitate genetic testing in FH patients. Methods and results: We designed a custom DNA resequencing array to detect mutations on all 3 FH-causing genes - LDL receptor (LDLR), apolipoprotein B (APOB), and proprotein convertase subtilisin/kexin type 9 gene (PCSK9) - and 290 known insertion/deletion mutations on LDLR. We verified FH array performance by analyzing 35 previously sequenced subjects (21 with point mutations, 2 insertions, 7 deletions, and 5 healthy controls) and blindly screening 125 FH patients. The average microarray call rate was 98.45{\%} and the agreement between microarray and capillary sequencing was 99.99{\%}. The FH array detected mutations by using automated software analysis, followed by manual review in 28 of the 30 subjects (pickup rate, 93.3{\%}). In the blinded study, the FH array detected at least 1 mutation in 77.5{\%} of patients clinically diagnosed with definite FH according to Simon Broome FH criteria and in 52.9{\%} with probable FH diagnosis. Conclusions: The high-throughput FH resequencing array detects LDLR, APOB, and PCSK9 with high efficiency and accuracy and identifies disease-causing mutations. Thus, it facilitates large-scale screening of the heterogeneous FH populations.",
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