High-resolution melting analysis for detection of familial ligand-defective apolipoprotein B-100 mutations

Ann Clin Biochem 2008;45:170-176
doi:10.1258/acb.2007.007077
© 2008 Association for Clinical Biochemistry

This Article
Right arrow
Figures Only
Right arrow
Full Text
Right arrow

Full Text (PDF)

Right arrow
Alert me when this article is cited
Right arrow
Alert me if a correction is posted
Services
Right arrow
Email this article to a friend
Right arrow

Similar articles in this journal

Right arrow
Similar articles in PubMed
Right arrow
Alert me to new issues of the journal
Right arrow
Download to citation manager
Right arrow
Citing Articles
Right arrow Citing Articles via Google Scholar
Google Scholar
Right arrow
Articles by Liyanage, K. E
Right arrow
Articles by van Bockxmeer, F. M
Right arrow Search for Related Content
PubMed
Right arrow
PubMed Citation
Social Bookmarking

What’s this?

Original Article


Khemanganee E Liyanage 1 *,
Amanda J Hooper 2 *,
Joep C Defesche 3,
John R Burnett 2 4 and
Frank M van Bockxmeer 1 2 


1 School of Surgery and Pathology, University of Western Australia, Perth, Australia;
2 Department of Core Clinical Pathology and Biochemistry, PathWest Laboratory Medicine WA, Royal Perth Hospital, Perth, Australia;
3 Department of Vascular Medicine, Academic Medical Centre, University of Amsterdam, Amsterdam, The Netherlands;
4 School of Medicine and Pharmacology, University of Western Australia, Perth, Australia


Corresponding author: Associate Prof Frank van Bockxmeer. Email: Frank.VB{at}health.wa.gov.au


Background: Familial ligand-defective apolipoprotein B-100 (FDB) is characterizedby elevated plasma concentrations of LDL-cholesterol and apolipoprotein(apo) B, normal triglyceride and HDL-cholesterol levels, thepresence of tendon xanthomas, and premature coronary arterydisease. FDB cannot be clinically distinguished from heterozygousLDL-receptor-defective familial hypercholesterolaemia (FH) withoutgenetic testing.

Methods: Amplicons in exon 26 and exon 29 of the APOB gene were screenedfor established genetic variants including mutations and polymorphismsusing high-resolution melting analysis. Six novel variants associatedwith FDB in hypercholesterolaemic Dutch patients (S3476L, S3488G,Y3533C, T3540M, I4350T, G4368D) were also studied.

Results: All positive controls, a total of 10 mutations in exon 26 andfour mutations in exon 29, were readily detectable by meltingcurve analysis. In addition, a patient previously not knownto be heterozygous for the H3543Y mutation was identified ina screen of hypercholesterolaemic subjects. The method was validatedby comparison of high-resolution melting analysis with DNA sequencedata in a ‘blinded’ manner in 35 consecutive patientsattending a lipid disorders clinic. These patients were classifiedas ‘definite FH’ by the Dutch Lipid Clinic Networkcriteria. Five patients were found to be heterozygous for theR3500Q and one for H3543Y.

Conclusions: We have established a novel, robust method of FDB mutation detectionusing high-resolution melting analysis in conjunction with DNAsequencing. Compared with existing methods it is not only morecost-effective, but is also capable of detecting new sequencechanges and will have importance in cascade screening of affectedsubjects.


CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What’s this?