Use of denaturing HPLC to provide efficient detection of mutations causing familial hypercholesterolemia

Olaf A. Bodamer, Dan Bercovich, Michael Schlabach, Christie Ballantyne, Danièle Zoch, Arthur L. Beaudet

Research output: Contribution to journalArticle

25 Citations (Scopus)

Abstract

Background: Autosomal dominant familial hypercholesterolemia (FH) attributable to mutations in the LDL receptor (LDLR) gene is one of the most common genetic disorders associated with significant morbidity and mortality. Definitive diagnosis would help to initiate appropriate treatment to prevent premature cardiovascular disease. Currently, clinical diagnosis of FH is imprecise, and molecular diagnosis is labor-intensive and expensive because of the size of the LDLR gene and number of coding exons. Methods: We used PCR to amplify all exons, including exon/intron boundaries, and the promoter of the LDLR gene. Nine individuals from five families with typical findings for a clinical diagnosis of heterozygous FH, 2 heterozygous FH cell lines, and 50 control individuals were screened for mutations by denaturing HPLC (DHPLC) followed by direct sequencing of aberrantly migrating fragments. Results: Mutations that were previously reported to be disease causing were identified in eight of nine individuals with FH and both cell lines (V502M, C146X, E207X, C660X, C646Y, and delG197), but none were found in controls. The one individual with FH in whom no mutation was found had a previously unreported change in the 5′-untranslated region of unknown significance. In addition, we identified several previously reported polymorphism both in controls and individuals with FH. Conclusions: DHPLC can be used to detect mutations causing FH. On the basis of our current experience with DHPLC, this method combined with confirmatory DNA sequencing is likely to be sensitive and efficient.

Original languageEnglish
Pages (from-to)1913-1918
Number of pages6
JournalClinical Chemistry
Volume48
Issue number11
StatePublished - Nov 1 2002

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Hyperlipoproteinemia Type II
LDL Receptors
High Pressure Liquid Chromatography
Exons
Mutation
Genes
Cells
5' Untranslated Regions
Polymorphism
Introns
Personnel
Cell Line
DNA
Inborn Genetic Diseases
DNA Sequence Analysis
Cardiovascular Diseases
Morbidity
Polymerase Chain Reaction
Mortality

ASJC Scopus subject areas

  • Clinical Biochemistry

Cite this

Bodamer, O. A., Bercovich, D., Schlabach, M., Ballantyne, C., Zoch, D., & Beaudet, A. L. (2002). Use of denaturing HPLC to provide efficient detection of mutations causing familial hypercholesterolemia. Clinical Chemistry, 48(11), 1913-1918.

Use of denaturing HPLC to provide efficient detection of mutations causing familial hypercholesterolemia. / Bodamer, Olaf A.; Bercovich, Dan; Schlabach, Michael; Ballantyne, Christie; Zoch, Danièle; Beaudet, Arthur L.

In: Clinical Chemistry, Vol. 48, No. 11, 01.11.2002, p. 1913-1918.

Research output: Contribution to journalArticle

Bodamer, OA, Bercovich, D, Schlabach, M, Ballantyne, C, Zoch, D & Beaudet, AL 2002, 'Use of denaturing HPLC to provide efficient detection of mutations causing familial hypercholesterolemia', Clinical Chemistry, vol. 48, no. 11, pp. 1913-1918.
Bodamer OA, Bercovich D, Schlabach M, Ballantyne C, Zoch D, Beaudet AL. Use of denaturing HPLC to provide efficient detection of mutations causing familial hypercholesterolemia. Clinical Chemistry. 2002 Nov 1;48(11):1913-1918.
Bodamer, Olaf A. ; Bercovich, Dan ; Schlabach, Michael ; Ballantyne, Christie ; Zoch, Danièle ; Beaudet, Arthur L. / Use of denaturing HPLC to provide efficient detection of mutations causing familial hypercholesterolemia. In: Clinical Chemistry. 2002 ; Vol. 48, No. 11. pp. 1913-1918.
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