Medizinische Universität Graz Austria/Österreich - Forschungsportal - Medical University of Graz
Gewählte Publikation:
Nauck, M; Wieland, H; März, W.
Evaluation of the Roche Diagnostics LightCycler-Apo B 3500 Mutation Detection Kit.
Clin Chem Lab Med. 2000; 38(7):667-671
Doi: 10.1515/CCLM.2000.096
Web of Science
PubMed
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- Co-Autor*innen der Med Uni Graz
- März Winfried
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- Abstract:
- Familial defective apolipoprotein (apo) B-100 is an autosomal codominant disorder associated with hypercholesterolemia and an increased risk of coronary artery disease. Two independent mutations affecting the codon 3500 (Arg3500-->Gln and Arg3500-->Trp) have been shown to cause ligand-defective apo B-100. Identification of carriers of these mutations is an important step in the risk stratification of individuals and families with hypercholesterolemia. We evaluated a homogeneous assay for detection of mutations at codon 3500 that combines rapid-cycle PCR with allele-specific fluorescent probe melting profiles for product genotyping. This single-tube analysis is performed on the LightCycler, a microvolume fluorimeter integrated with a thermal cycler. Continuous acquisition of fluorescence data during a melting curve analysis at completion of PCR allows the detection of mutations, as loss of fluorescence occurs in an allele-specific manner. By plotting melting peaks, the three apo B-100 alleles were readily distinguishable. Using this method, genotyping of 32 samples is completed within 40 min without the need for any post-PCR sample manipulation, thereby eliminating the risks of end-product contamination and sample tracking errors. The specific detection of mutations at codon 3500 of the apo B gene on the LightCycler is a rapid and reliable method that is ideally suitable for typing both small and large numbers of samples.
- Find related publications in this database (using NLM MeSH Indexing)
- Apolipoprotein B-100 -
- Apolipoproteins B - genetics
- DNA - analysis
- Fluorescence - analysis
- Genotype - analysis
- Humans - analysis
- Hyperlipoproteinemia Type II - genetics
- Point Mutation - genetics
- Polymerase Chain Reaction - economics
- Reagent Kits, Diagnostic - economics
- Reproducibility of Results - economics
- Sensitivity and Specificity - economics
- Find related publications in this database (Keywords)
- apo B-100
- familial defective apo B
- PCR
- hybridization probes
- fluorescence resonance energy transfer
- hypercholesterolemia