Apolipoprotein E haplotyping by denaturing high-performance liquid chromatography
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Maura Poli
Abstract
The apolipoprotein E ( APOE) gene in humans contains two single-base polymorphisms in exon 4, which result in three common alleles, conventionally named ε2, ε3 and ε4. Numerous studies have shown an important association between the ε4 variant and an increased risk of Alzheimer's disease; other data suggest a possible linkage of APOE genetic heterogeneity with lipid profile and an increased risk of atherothrombotic stroke and coronary heart disease. APOE genotyping is therefore an increasingly common assay in laboratory medicine. The most widely used technique for APOE genotyping is based upon restriction isotyping, i.e., amplification of the fragment of exon 4 containing the most common se-quence variations, followed by enzymatic digestion of the amplicon and fragments analysis by gel electrophoresis. We developed a novel, reliable and fast method that exploits the sensitivity and specificity of denaturing high-performance liquid chromatography in detecting single-nucleotide polymorphisms. We show that, in most cases, with a single chromatographic separation it is possible to correctly identify the six different APOE allelic patterns, without any manipulation after the polymerase chain reaction amplification. When compared to restriction isotyping, our method is much faster, less labor intensive and similarly inexpensive.
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© by Walter de Gruyter Berlin New York
Articles in the same Issue
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- Apolipoprotein E haplotyping by denaturing high-performance liquid chromatography
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Articles in the same Issue
- Second Santorini Conference “From Human Genetic Variations to Prediction of Risks and Responses to Drugs and to the Environment”
- Expressed genome molecular signatures of heart failure
- Understanding hyperlipidemia and atherosclerosis: lessons from genetically modified apoe and ldlr mice
- Alcohol and gene interactions
- β-Carotene stimulates chemotaxis of human endothelial progenitor cells
- Effect of interferon-γ, interleukin-10, lipopolysaccharide and tumor necrosis factor-α on chitotriosidase synthesis in human macrophages
- Two immunochemical assays to measure advanced glycation end-products in serum from dialysis patients
- Apolipoprotein E haplotyping by denaturing high-performance liquid chromatography
- Both core and terminal glycosylation alter epitope expression in thyrotropin and introduce discordances in hormone measurements
- Do we measure bilirubin correctly anno 2005?
- Differences in mortality on the basis of laboratory parameters in an unselected population at the Emergency Department
- An Italian program of external quality control for quantitative assays based on real-time PCR with Taq-Man™ probes
- A reference material for traceability of aspartate aminotransferase (AST) results
- Harmonization of the Bayer ADVIA Centaur and Abbott AxSYM automated B-type natriuretic peptide assay in patients on hemodialysis
- Multicenter evaluation of the analytical and clinical performance of the Elecsys ® S100 immunoassay in patients with malignant melanoma
- Guidelines for sampling, measuring and reporting ionized magnesium in undiluted serum, plasma or blood: International Federation of Clinical Chemistry and Laboratory Medicine (IFCC): IFCC Scientific Division, Committee on Point of Care Testing
- Evaluation of the Quantase™ neonatal immunoreactive trypsinogen (IRT) screening assay for cystic fibrosis
