Integrated single-label liquid-phase assay of APOE codons 112 and 158 and a lipoprotein study in British women

Clinical Chemistry
Mohammad Reza AbdollahiIan N M Day

Abstract

Apolipoprotein E (APOE) is an important element of lipid metabolism and, hence, cardiovascular disorders. APOE has 3 main allelic variants: epsilon3, epsilon4, and epsilon2. Of these, epsilon3 is the most common, followed by epsilon4 and epsilon2. The associations of these isoforms with cardiovascular disorders and Alzheimer disease have been widely studied in different populations. Most of the genotyping in these studies has been performed with gel-based methods, which have important limitations, particularly for large epidemiologic studies. We therefore developed an integrated "one-tube" liquid-phase assay. To measure APOE isoforms, we developed an integrated single-label liquid-phase fluorescence assay containing 2 PCR primers, 2 probes, and 2 quencher oligonucleotides. We used a 384-well LightTyper, but the assay would be generically applicable for use with any fluorescence detector with thermal ramp control. We validated this method and applied it in the British Women's Heart and Health Study. There were 4 melting peaks, at 41, 56, 61, and 69 degrees C, which generated 6 distinctive patterns representing genotypic combinations of epsilon3, epsilon4, and epsilon2. The magnitude and direction of the associations found with t...Continue Reading

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Citations

Oct 10, 2008·Clinical and Experimental Medicine·B H RihnA Le Faou
Jun 23, 2012·Laterality·Brian J PiperJeremy K Miller
Aug 20, 2015·PloS One·Lindsey I SinclairGlyn Lewis
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May 8, 2021·Clinical Medicine Insights. Case Reports·Marina YukinaAlina Bratchikova

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