Maura Poli, Luisa Benerini Gatta, Roberto Dominici, Carlo Lovati, Claudio Mariani, Alberto Albertini, Dario Finazzi
Apolipoprotein E haplotyping by denaturing high-performance liquid chromatography
The apolipoprotein E (
APOE) gene in humans contains two single-base polymorphisms in exon 4, which result in three common alleles, conventionally named
?2,
?3 and
?4. Numerous studies have shown an important association between the
?4 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.
Clinical Chemical Laboratory Medicine, Walter de Gruyter
Print ISSN: 1434-6621
Volume: 43, 05/2005
Pages: 512 - 518
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